Austin J Biosens & Bioelectron. 2015; 1(3): 1015.
Rodovalho VR1, Alves LM1, Castro ACH1, Madurro JM2, Brito-Madurro AG1 and Santos AR3*
1Institute of Genetics and Biochemistry, Federal University of Uberlandia, Brazil
2Institute of Chemistry, Federal University of Uberlandia, Brazil
3Faculty of Computing, Federal University of Uberlandia, Brazil
*Corresponding author: Santos AR, Faculty of Computing, Federal University of Uberlandia, Avenue Jo&aTilde;o Naves de Ávila 2121, Campus Santa Mônica, Block 1B, Room 120, 38400-902, Brazil
Received: September 26, 2015; Accepted: December 30, 2015; Published: December 31, 2015
In the past five decades, biosensors have consolidated their impact in several fields, including clinical applications, due to advantages such as high selectivity and sensitivity, potential for miniaturization, portability, low cost and rapid response. Recent advances in biomarkers discovery and biotechnology are now clarifying the nuances of many biological processes in health and disease, highlighting new targets for diagnosis and therapeutics. This is especially important in the case of infectious diseases, since the number of predicted deaths remains high, with threats of epidemics and pandemics, emerging and re-emerging diseases and pathogen resistance to antibiotics. Therefore, the availability of robust diagnosis methods is crucial. This review presents the current strategies for diagnosis of infectious diseases, notions about biomarkers and ligand selection, besides focusing on the promising technology of biosensors.
Keywords: Biosensors; Sensors; Biomarkers; Diagnosis; Infectious diseases
ELISA: Enzyme-Linked Immunosorbent Assay; PCR: Polymerase Chain Reaction; MALDI-TOF MS: Matrix-Assisted Laser Desorption/ Ionization Time-Of-Flight Mass Spectrometry; NGS: Next Generation Sequencing; FISH: Fluorescence In Situ Hybridization; SELEX: Exponential Enrichment; SPR: Surface Plasmon Resonance; WHO: World Health Organization; EIS: Electrochemical Impedance Spectroscopy
Infectious diseases are caused by pathogenic microorganisms, including bacteria, viruses, fungi and parasites. Some examples among those enumerated by the World Health Organization (WHO) include tuberculosis, meningococcal meningitis, malaria, AIDS, pneumonia, poliomyelitis, hepatitis, Ebola virus disease, dengue and Chikungunya, American trypanosomiasis (Chagas disease), leprosy, toxoplasmosis and leishmaniasis.
The urbanization process and the consequent lack of city planning, poor management of sanitary conditions and water supplies, great inhabitants’ density and interference in previously untouched ecosystems conjunctly contribute to the spread of infectious diseases . In spite of the existent vaccination programs, the rise in incidence of certain diseases shows the impact of intentional under vaccination and the urge for public health education programs , as well as the need of new immunization strategies and alternatives to overcome pathogen resistance to antibiotics .
Although the mortality related to infectious diseases is being reduced worldwide, the number of deaths predicted for 2050 is 13 million and the threats of epidemics and pandemics remain considerable . Moreover, emerging and re-emerging diseases caused by new, uncategorized or persistent pathogens have been reported .
This review presents the current strategies for diagnosis of some infectious diseases, notions about biomarkers and ligand selection, besides focusing on the promising technology of biosensors.
A diagnostic test is any method for identification of a patientâ€™s disease or condition. In the case of infectious diseases, it allows the detection of presence or absence of infection. The importance of simple, accurate, affordable and rapid diagnosis tests is justified by its impact in the clinical management, since early diagnosis affects therapy effectiveness and avoids long-term complications and pathogen transmission .
While the standard diagnosis techniques for infectious diseases include well-established methodologies, such as Enzyme- Linked Immunosorbent Assay (ELISA), nucleic acid-based assays, microscopy and microorganism culture , the development of new strategies for the evaluation of specific biomarkers in clinical diagnosis is imperative . The diagnosis tests for infectious diseases should present a set of desirable characteristics, such as sensitivity, specificity and reproducibility .
Historically, the identification of infectious agents was initially performed by culture and microscopy. Then, antigen detection and Polymerase Chain Reaction (PCR) became widely used. Currently, pathogen identification and host response (e.g., antibodies detection) are both used to diagnosis pathological states .
PCR and DNA microarrays are two widely used nucleic acids technologies. PCR employs oligonucleotide primers that are complementary to pathogen genetic material to amplify it, if present in the sample. The reaction product is detected during or after the process. Microarray technology, by the other hand, allows multiple target detection through hybridization with the probes immobilized on a surface .
Recently, Matrix-Assisted Laser Desorption/Ionization Time- Of-Flight Mass Spectrometry (MALDI-TOF MS) is being adopted in the comparison of protein fingerprint obtained in a sample with the available databases, for the identification of bacteria, fungi  and viral pathogens . There is also the potential of Next Generation Sequencing (NGS) methodologies to revolutionize infectious diseases diagnosis, since it does not rely on pre-established sequence targets, allowing the identification of emerging or mutating pathogens [12,13]. Other strategies are also being developed for clinical diagnosis, including microfluidic  and nanotechnological  devices.
Biosensors, other promising diagnosis technology that has received attention in the last decades and has several associated advantages will be discussed later in detail.
A biomarker is “a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention” . Biomarkers are also defined as specific variables, represented by biomolecules, such as genes, proteins and metabolites, associated to certain populations as distinguishable features . Moreover, this term may be associated with the use of genomics, transcriptomics, proteomics and metabolomics technologies, the monitoring of drug discovery, as well as clinical concepts, such as prediction, progression, regression, outcome, diagnosis and therapeutics . Some examples of biomarkers for infectious diseases include C-Reactive Protein (CRP) and soluble Triggering Receptor Expressed on Myeloid cells 1 (sTREM-1). While CRP is applied for sepsis, severe infection, rheumatologic conditions and coronary artery disease risk stratification; sTREM-1 is a sepsis prognostic marker .
The ideal biomarkers should present a whole set of desirable characteristics, including accessibility for measurement, sensitivity and specificity. Its clinical importance should be externally validated; its use should result in cost-effective assays  and present reproducibility and stability toward sample variations .
Biomarkers identification may be achieved by several approaches, including simple statistical tests, development and analysis of classification models or subset-selection optimization . In the last decades, several biomarkers for infectious diseases have been identified, due to improvements in biomolecules screening techniques and bioinformatics analysis. However, their translation into clinical use is still limited .
Depending on the class of biomarker, there are several methods for the selection of specific ligands, including phage display and Systematic Evolution of Ligands by Exponential Enrichment (SELEX) .
Polymerase Chain Reaction (PCR) and Fluorescence In Situ Hybridization (FISH) are two widely used nucleic acid detection technologies. While PCR consists in the in vitro specific DNA amplification, FISH is the fluorochrome-labeling of oligonucleotides for hybridization with the complementary target. Both methods rely on the use of known DNA sequences that relates to specific pathogens [22,23]. For the development of such diagnostic systems based on nucleic acid biomarkers and ligands, the general strategy is to compare sequences in order to find species-specific oligonucleotide targets that are present in high copy numbers in the cells .
When designing DNA oligonucleotide probes for several applications, including PCR and DNA hybridization assays, some criteria should be followed: assure their complementarity and specificity for the target, avoid competing secondary structures (dimers and hairpins) and choose adequate melting temperature [24,25].
Most efforts in nucleic acid probes design has been related to PCR , FISH  and microarray technologies . Cho et al.  reported the selection of 23-mer primers for Vibrio cholera detection by quantitative polymerase chain reaction. The primers pair had the outer membrane lipoprotein lolB gene as target and allowed the specific identification of several isolates. Ogura et al.  reported a method of microarray probe design based on comparison of edit distance between sequences to avoid cross hybridization with similar probes on the array. Naidoo et al.  evaluated Mycobacterium Tuberculosispili (MTP) gene and protein sequences as potential biomarkers for tuberculosis and found they are specific and highly conserved among strains of the Mycobacterium Tuberculosis Complex (MTBC), through BLAST and multi-sequence alignment.
These examples show that DNA-based diagnosis and selection of appropriate biomarkers ligands have focused in comparing sequences. However, this paradigm is changing with the application of Next-Generation Sequencing (NGS), which does not rely on previous knowledge about the pathogens genetic sequence .
Several techniques are employed for protein biomarker discovery and ligands selection, including mass spectrometry, gel electrophoresis and protein microarrays, the later allowing the study of entire proteomes . Also, peptides microarrays may be employed in the selection of peptide ligands, after computational preselection .
Another notable methodology is called phage display and relies on the expression and presentation of a great diversity of peptides or proteins on bacteriophages surfaces, allowing their selection against a target biomolecule . Therefore, this tool can be used to find new reagents for immunological assays, including phage-displayed peptides that mimetize pathogen antigens, with applications like leprosy diagnosis  and development of vaccines for visceral leishmaniasis . Wu et al.  selected three single-chain variable Fragments (scFv) for detection of Highly Pathogenic Avian Influenza A (HPAI) viruses strains, which could be important to accelerate diagnosis and control outbreaks. Another application is the presentation of antibodies in the bacteriophage surface for potential therapeutic purposes , with targets such as the CCR5 HIV coreceptor .
Aptamers are high-affinity ligands selected in vitro. Their targets may be specific proteins isoforms or conformations, being analogous to antibodies . Aptamersare selected from libraries through Systematic Evolution of Ligands by Exponential enrichment (SELEX) against several classes of targets, including carbohydrates, proteins and inorganic molecules .
Aimaiti et al.  selected species-specific aptamers for discrimination of Mycobacterium tuberculosis strains through whole cell SELEX process. Shiratori et al.  developed DNA aptamers for proteins of several influenza A virus subtypes and applied them in a successful sandwich detection method. Besides diagnostic applications, aptamers have also been used as therapeutics agents, such as the recently reported S15 aptamer, that binds to the envelope protein of all dengue serotypes, neutralizing the infections .
These examples show how the integration of physiology, biochemistry, genetics, bioinformatics and other research fields, may improve biomarker discovery and selection of specific ligands, allowing a better understanding of infectious diseases and the development of effective diagnostic systems, such as biosensors .
Biosensors are analytical devices that convert a biochemical recognition event into a measurable signal , consisting mainly of a probe (biological recognition element) and a physicochemical detector (transducer) (Figure 1). The objective is to determine the presence, activity or concentration of an analyte in a solution , with a broad range of applications, such as industrial , environmental  and medical [49,50].
Figure 1: Schematic representation of the general operation principle of biosensors. The biological element is immobilized on a surface. In the presence of a specific target analyte that binds to the immobilized biomolecule, the transducer converts the recognition event into a measurable signal, which can be then processed (Adapted from Ronkainen et al ).
Biosensors present several advantages over other analytical tools, such as high selectivity and sensitivity, potential for miniaturization and portability , low cost, detection in real time, use of small sample volumes and rapid response [51-53].
Depending on specific criteria, biosensors may be classified into distinct groups. Regarding the transducer type, there are the calorimetric sensors, related to the conversion of enthalpy , the piezoelectric, sensitive to mass changes , the optical, based on light-associated phenomena  and the electrochemical, based on the generation or consumption of electro active species .
Another classification system is based on the biological component (probe), which can be DNA molecules , enzymes , and antibodies , among others (Figure 2). Enzymatic biosensors take advantage of the catalytic activity, selectivity and specificity of enzymes. The pioneer works of Clark et al. , such as the coupling of the enzyme glucose oxidase to an electrode that recognized the oxygen uptake, yielded one of the most successful classes of biosensors. By the other hand, genosensors contain DNA fragments immobilized on their surface and can detect mutated genes associated with human diseases , as well as the genetic identification of pathogens [62- 64]. Immunosensors contain antibodies as biological element, whose specific binding sites interact with the antigen to form an antibodyantigen complex . Other types of biologic elements for biosensors include aptamers , cells and tissues , and microorganisms .
Figure 2: Schematic representation of distinct groups of biosensors, regarding the biological component. The surface-immobilized biomolecules glucose oxidase (PDB1GPE ), anti-HIV-1 antibody (PDB 1HZH ) and a DNA dodecamer (PDB 1BNA ) represent the enzymatic biosensors, immunosensors and genosensors, respectively. Other types of biologic elements for biosensors (not represented) include aptamers, cells and tissues, and microorganisms. The arrows indicate that, as a general principle, the binding of a specific target is converted into a measurable signal.
Although different types of biosensors have been successfully developed and applied to the medical field, for the diagnosis of pathologies such as cancer [69-72], cardiovascular [73-75], autoimmune [76-78] and neurodegenerative diseases [79-82], this review will focus on infectious diseases.
Viruses are infectious agents that may be responsible for several diseases in humans, including Human Papilloma Virus (HPV)  (Table 1), dengue virus [43,84-86] and hepatitis virus [62,87-89].
Biomarker / probe
n.s. / DNA oligonucleotide
1 x 10-8 - 1 x 10-13 M
4.03 x 10-14 M
n.s. / 5051 mAb
PMMA/gold nanolayer/4-ATP SAM
n.s. / DNA oligonucleotide
12.5 â€“ 350.0 nM
Hepatitis B virus
n.s. / DNA oligonucleotide
1.89 x 10-9 - 1.89 x 10-6 M
Hepatitis B virus
HBsAg / mAb
1.7 - 1920 pg mL-1
0.358 pg mL-1
n.s. / DNA oligonucleotides
NS1 protein / anti-NS1 antibody
0.01â€“2.00Â Âµg mL-1 (PBS)
0.01â€“1.00Â Âµg mL-1 (Neat blood)
3 ng mLâˆ’1 (PBS)
30 ng mLâˆ’1 (Neat blood)
NS1 protein / anti-NS1 antibody
0.04 â€“ 0.6Â Âµg mL-1
0.015 Âµg mL-1
n.s.: not specified; 4-ATP: 4-Aminothiophenol; AuNpPANI: Gold Nanoparticles-Polyaniline Hybrid Composite; G: Grapheme; AuNRs: Gold Nanorods; GCE: Glassy Carbon Electrode; GE: Gold Electrode; GrE: Graphite Electrode; HBsAg: Hepatitis B Surface Antigen; HPV: Human Papillomavirus; mAb: monoclonal Antibody; MUA: 11-Mercaptoundecanoic Acid; PMMA: Polymethylmethacrylate; PT: Polythionine; SAM: Self-Assembled Monolayer; SPE: Screen-Printed Electrode.
Table 1: Characteristics from several biosensors for detection of pathogenic virus.
There are 100 genotypes of HPV virus and some of them are associated with cancer, especially in the cervix and anus . The methods used for the diagnosis have limitations, such as low specificity . To overcome this disadvantage, Huang and coworkers described a highly sensitive electrochemical biosensor based on DNA probes for Human Papillomavirus (HPV), using a glassy carbon electrode functionalized with graphene, gold nanorods and polymeric film. They used electrochemical impedance spectroscopy and 1,10-phenanthroline ruthenium dichloride (Ru(phen)3 2+)as redox indicator, amplifying the electrochemical signal. The biosensor described proved to be efficient in the viral DNA detection, specifically detecting the target in human serum samples with a detection limit of 4.03 x 10-14 M .
Urrego and collaborators described a biosensor for the detection of HPV that was able to perform 98 simultaneous tests. It was based on a monolayer of 4-aminophenol on a surface of poly(methyl methacrylate) with a gold nanolayer and the immobilization of a monoclonal antibody (mAb 5051) specific for HPV 16, one of the most common genotypes among women. Electrochemical impedance spectroscopy was used in the analysis. The biomicrosystem developed was portable, used a small volume of sample and simple equipment .
Nasirizadeh et al developed a genosensor using gold electrodes, thiolated oligonucleotides specific for HPV and monitored the interaction of hematoxylin with dsDNA formed after the hybridization process. They used the techniques of cyclic voltammetry and differential pulse voltammetry, observing a remarkable difference between the voltammetric signals in different samples after hybridization. The linear relationship with the concentration of DNA target varied from 12.5 to 350.0 nM and the detection limit was 3.8 nM .
Another infectious disease caused by a DNA virus that infects hepatocytes of the liver is hepatitis B. It is a global health problem with approximately 2 billion people infected, corresponding to about a third of the world having positive serology for hepatitis B . Hepatitis B virus infection can harm the liver, with high risk of death from liver cirrhosis and cancer . During the chronic phase of the disease, monitoring is crucial, since it prevents the development of progressive diseases, such as cirrhosis and liver failure, as well as hepatocellular carcinoma.
As an alternative to the traditional methods, Castro and coworkers developed an electrochemical biosensor for the detection of a specific DNA sequence of the hepatitis B virus, using graphite electrodes modified with poly (4-aminophenol), differential pulse voltammetry as detection technique and ethidium bromide as hybridization label (Figure 3). They showed that this device was effective for diagnosis in the serum of infected patients and had a detection limit of 2.61 nM .
Figure 3: Example of genossensor for detection of a specific DNA sequence before (left) and after (right) the binding of the target (Adapted from Castro et al ). A specific DNA oligonucleotide was immobilized on a graphite/ poly (4-aminophenol) surface and it was blocked with BSA. Then, the complementary DNA target was applied. Ethidium bromide was used to discriminate the single-stranded and double-stranded DNA.
Shourian et al.  developed a colorimetric immunosensor to detect the surface antigen of hepatitis B virus. The strategy used was a sandwich immunoassay system that had gold nanoparticles functionalized with biotin and luminol. A chemiluminescent signal was produced by the gold nanoparticles in the presence of a catalyst and hydrogen peroxide as the oxidant. The immunosensor had a linear range of 1.7 to1920 pg mL-1 and the detection limit of 0.358 pg mL-1.
Dengue is a major public health problem in the world, with 2.5 billion people at risk of contracting the disease. It is transmitted by infected female mosquitoes Aedesaegypti and Aedesalbopictus, being considered endemic in 98 tropical and subtropical countries.
Current methods for diagnosis of dengue are based on the detection of viral RNA by Reverse Transcription Polymerase Chain Reaction (RT-PCR) or immunoassay methods including ELISA, where the analyte targets are antibodies raised in response to the viruses in the infected patient. However, early diagnosis of disease with these methods can be expensive, time-consuming, and not sensitive and produce false positives [85,93].
Trying to overcome these problems, Nascimento and coworkers described the development of a biosensor based on gold electrodes functionalized with a composite of gold nanoparticles, polyaniline and with SH-terminal groups (AuNpPANI-SH). On this surface, they immobilized 3 specific oligonucleotides for serotypes 1 (T1), 2 (T2) and 3 (T3) and identified the genomic material, using ferricyanide/ ferrocyanide potassium as indicator and cyclic voltammetry and electrochemical impedance spectroscopy as detection techniques. The system AuNpPANI-ST exhibited a highly selective response to the genome of dengue fever in human patients and can be used for the construction of a biosensor for serotypes of dengue in low concentration .
Cecchetto and coworkers decided to use the Non-Structural protein of dengue (NS1) as target for a biosensor. The authors developed an immunosensor using gold electrodes modified with a self-assembled monolayer of mercaptoundecanoic acid and immobilization of anti-NS1 antibody by covalent bonds. The detection technique was electrochemical impedance spectroscopy and the calibration curve linearity ranged between 0.01â€“2.00μg mL-1 in PBS and 0.01â€“1.00 μg mL-1 in serum, with a detection limit of 3ng mL-1 in PBS and 30ng mL-1 in serum .
Silva et al.  developed a screen-printed electrode using a modified graphite ink with thiophene and a layer of gold nanoparticles, immobilized anti-NS1 antibodies and detected the amperometric responses of the NS1 protein of the dengue virus by cyclic voltammetry in the presence of ferrocyanide/ferricyanide.
Linearity was obtained from 0.04 to 0.6 μg mL-1 and the detection limit was 0.015 μg mL-1.
Pathogenic bacteria are important targets for detection in several fields, such as medicine and food safety. Different approaches have been developed for the detection of pathogenic bacteria, since these microorganisms contribute to globally important diseases, such as tuberculosis, leprosy and meningitis [96-100] (Table 2).
Biomarker / probe
IS6110 gene / DNA oligonucleotide
1 x 10-15 - 1 x 10-9M
Genomic DNA / DNA oligonucleotide
GE/ MBA SAM/Fe3O4Nps
6 - 40Â ng ÂµL-1
6 ng ÂµL-1
n.s. / DNA oligonucleotide
1 x 10-14 - 1 x 10-9 M
8,7 x 10-15 M
IS6110 gene / DNA oligonucleotide
up to 100 aM
rpoB gene / DNA oligonucleotide
IS6110, 16S ribosomal RNA, 85B, Rv3130c and Rv3133c / DNA oligonucleotide
Mycobacterium tuberculosis, Mycobacterium avium
ITS gene / DNA oligonucleotides
104 CFU mL-1 - 108 CFU mL-1
4.2 x 104 CFU mL-1 and 3.7 x 104 CFU mL-1, respectively
Serum antibodies / Antigens (W06,W10,W14,W19,W28,W38,W64,W70,W85)
Array chip/8-MOA SAM
Ag85, ESAT6 and LAM / specific antibodies
Waveguides functionalized with a lipid bilayer
0.5 / 100 / 1 pM,
200 CFU mL-1
n.s. / mAb
100 CFU mL-1
n.s. / PCR product
0.35-35.0 ng ÂµL-1
IgM and IgG antibodies / ND-O and LID-1 antigens
Membrane protein 85 (Omp85) / specific antibody
0.3-20 Âµg mL-1
Omp85 gene/ DNA oligonucleotide
6-100 ng/6 ÂµL
6 ng/6 Âµl
CtrA gene / DNA oligonucleotide
7â€“42 ng ÂµL-1
CtrA gene / DNA oligonucleotide
10â€“60 ng ÂµL-1
CtrA gene/ DNA oligonucleotide
5â€“240 ng ÂµL-1
about 5 ng ÂµL-1
rmpM gene / DNA oligonucleotide
1-12 ng/6 ÂµL
3 ng/6 Î¼L
n.s.: not specified; 8-MOA: 8-Mercaptooctanoic Acid; AuNPs: Gold Nanoparticles; Fe3O4Nps: magnetite Nanoparticles; GCE: Glassy Carbon Electrode; GE: Gold Electrode; GrE: Graphite Electrode; mAb: monoclonal Antibody; MBA: Mercaptobenzoic Acid; Pt/Si: Platinum/Silicon; PVDF: Polyvinylidenedifluoride; rGO: reduced Graphene Oxide; SAM: Self-Assembled Monolayer; SPCE: Screen-Printed Carbon Electrode; SPGE: Screen-Printed Gold Electrode; ZNF: ZnOnanoflowers.
Table 2: Characteristics from several biosensors for detection of pathogenic bacteria.
Tuberculosis is caused by the pathogenic bacteria Mycobacterium tuberculosis and is currently the leading infectious cause of death, undoubtedly representing a global public health priority . According to the World Health Organization, in 2013 approximately 5.7 million cases of tuberculosis were reported worldwide.
In recent years, many biosensors platforms have been developed for tuberculosis based on different biological recognition elements and various transducers. Liu et al.  developed an electrochemical genosensor for M. tuberculosis based on the immobilization of a specific sequence of the IS6110 gene using a reduced graphene oxidegold nanoparticle-modified electrode as a sensing platform and gold nanoparticlesâ€“polyaniline as a tracer label for amplification.
The linear response of the sensor was 1 x 10-15 to 1 x 10-9M. Costa et al.  described an electrochemical genosensor based on selfassembled monolayers of mercaptobenzoic acid and magnetite nanoparticles (Fe3O4Nps) on bare gold electrode for immobilization of a DNA probe. The detection limit was 6ng μL-1.
Zhang et al.  proposed a novel electrochemical biosensing platform using CdSe quantum dots as a label combined with MspI endonuclease and gold nanoparticles to improve the selectivity and amplify the signal. The sensor linear range of response was 1 x 10-14 to 1 x 10-9 M and it discriminated mismatched DNA for M. tuberculosis with high selectivity. For the detection of very small quantities of pathogen genomic DNA, an electrochemical method was developed and this platform was applied to the detection of M. tuberculosis in sputum, pleural fluid and urine samples. This methodology is based on the entrapment of amplified single-stranded DNA sequences on magnetic beads, followed by the post-amplification hybridization assay to provide a higher level of specificity .
Other detection systems have been reported, such as Surface Plasmon Resonance (SPR), an optical detection technique that has been widely used for the development of genosensors for M. tuberculosis [105-107], and immunoassays [108-111].
Another disease caused by bacteria of the genus Mycobacterium is leprosy, a chronic disease caused by Mycobacterium leprae. According to WHO, in 2013 about 215,000 cases of leprosy were reported in the world, and the early diagnosis is important to interrupt transmission and prevent severe damage to patients .
Afonso et al.  developed an electrochemical genosensor based on the immobilization of a specific single-stranded DNA oligonucleotide on a graphite electrode modified with poly (4-aminophenol). The system target was M. leprae and the linear range of detection was from 0.35 to 35.0 ng μL-1. In addition, a fast and quantitative test for leprosy was developed by immobilizing two specific antigens on nitrocellulose membranes to detect IgMandIgG antibodies .
Meningitis can be caused by various pathogens, such as bacteria, fungi, viruses and parasites. Among the bacteria species that can cause meningitis, the most common are Streptococcus pneumoniae, Neisseria meningitidis, Haemophilusinfluenzae, Listeria monocytogenes and Streptococcus [115,116]. Particularly, Neisseria meningitidis has the potential to cause large epidemics.
Among the studied targets for detection, there is Omp85, a virulence gene that codes for a conserved outer membrane protein of N. meningitidis. Reddy et al.  described the development of an immunosensor using the quartz crystal microbalance as transducer and antibodies against the cell surface outer membrane protein 85 of N. meningitides as biologicalrecognition element (Figure 4). In addition, an electrochemical genosensor was developed using specific oligonucleotides for this virulent gene immobilized on screen-printed gold electrodes and the sensor sensitivity was 2.6 (μA/cm2)/ng .
Figure 4: Example of immunosensor for detection of meningococcal antigen before (left) and after (right) the binding of the target (Adapted from Reddy et al. ). A gold electrode was modified with polyvinylidenedifluoride thin film deposition. Antibodies were directionally orientated by interaction with protein A and the surface was blocked with casein. Next, gold nanoparticles conjugated with the target antigen and BSA was applied to the surface, in order to allow the antigen-antibody interaction.v
Other electrochemical genosensors have been described using specific oligonucleotides for other virulent factors. A genosensor was developed through the immobilization of thiol-labeled DNA probe on a gold electrode surface and its hybridization with complementary sequence of the ctrA gene. The sensitivity was 115.8 μA/ng with 0.917 regression coefficient , 0.0115 μA/ng with 0.999 regression coefficient  and 168,64 μA/ng with 0.98 as regression coefficient . Dash et al.  used specific oligonucleotides complementary to the rmpM (reduction-modifiable protein M) virulent gene as a biological recognition element. The sensitivity of the genosensor was 9.5087 μA/ng.
Protozoa are one of the main classes of parasites that cause diseases in humans. A wide variety of approaches have been applied to the development of biosensors for the diagnosis of protozoan-caused diseases such as malaria, leishmaniasis, American trypanosomiasis (Chagas disease) and toxoplasmosis.
Malaria is transmitted to humans by the bite of more than thirty species of female anopheline mosquitoes. The etiologic agent is a protozoan of genus Plasmodium. Five species, P. falciparum, P. vivax, P. malariae, P. ovale, and P. knowlesi, are known to affect humans . According to the WHO, in 2013 about 48 million cases of malaria were reported worldwide, with 584,000 deaths (Table 3).
Biomarker / probe
Anti-PfHRP-2 antibodies/ PfHRP-2
SPE/Al2O3 solâ€“gel/ AuNPs
1 : 102400 dilution of rabbit anti-PfHRP-2
PfHRP-2 / specificantibody
8 ng mL-1
PfHRP-2 / specificantibody
Anti-PfHRP-2 antibodies / PfHRP-2
5.6 pgand 0.4 ng
Plasmodium falciparum and Plasmodiumvivax
pLDH / aptamer
1- 1000 pM
pLDH / aptamer
AuNPs/PDDA or PAH
74 parasites/Âµl (PAH)
Plasmodium falciparum and Plasmodium vivax
pLDH / aptamer
1 pM - 1 nM
1.25 pM (Plasmodiumvivax)and 2.94 pM (Plasmodiumfalciparum)
Plasmodium falciparum and Plasmodium vivax
n.s. / DNA oligonucleotide
antibodies / rK39 + LicTXNPxantigens
antibodies / L. infantumantigens
Parasite antigens / mABs
1.8 x 10-4 amastigotes/g of infected tissue
18S rRNA gene / DNA oligonucleotide
2pg/ml - 2Î¼g/ml
0.02 Â±0.002 ng/Î¼l
T. cruziantibodies / CRA and FRA antigens
GE and PE
T. cruziantibodies / T. cruziantigen
IgG antibodies / specific antigens
11 - 205 ng mL-1
3.065 ng mL-1
IgG antibodies / specific antigen
PQC/ n-butyl amine PPF
IgG antibodies / specific antigen
IgG antibodies / aptamer
n.s. / DNA oligonucleotides
n.s.: not specified; 4-MBA:4-Mercaptobenzoic Acid; Al2O3solâ€“gel: Alumina solâ€“gel; AuNPs: gold Nanoparticles; CA: Cysteamine; CRA: Cytoplasmic Repetitive Antigen; DDT: 1-Dodecanethiol; Fe3O4/CdTe: magnetic fluorescent nanoparticles; FRA: Flagellar Repetitive Antigen; GE: Gold Electrode; ITO: Indium Tin Oxide; MPA: 3-Mercaptopropionic Acid; MUA: 11-Mercaptoundecanoic; MWCNT: Multiwall Carbon Nanotubes; NiO: Sol-gel synthesized nickel Oxide; PAH: Poly (Allylamine
Table 3: Characteristics from several biosensors for detection of pathogenic bacteria.
Various biomarkers have been used to malaria diagnosis . Histidine-rich protein-II is produced and secreted by the parasite during its growth and development and it has been widely used for the development of electrochemical [123-125] and optical immunosensors . Biosensors based on the immobilization of aptamers with high affinity for lactate dehydrogenase, another biomarker for malaria, has been reported in the literature using electrochemical  and colorimetric [128,129] transducers. Reddy et al.  describes the development of Plasmodium lactate dehydrogenase-specific ssDNA aptamers by SELEX using magnetic beads. The selected aptamers were characterized and used for the construction of an aptamerbased electrochemical sensor able to discriminate malaria positive samples from non-infected sample (Figure 5). In addition, Ittarat et al.  described a genosensor based on quartz crystal microbalance to differentially diagnose malaria infection by either P. falciparum or P. vivax.
Figure 5: Example of aptasensor for the diagnosis of malaria before (left) and after (right) the binding of the target (Adapted from Lee et al. ). The scheme illustrates a specific thiol-modified aptamer immobilized on a gold electrode, together with spacer molecules. Next, the protein target was applied for interaction with the aptamer probe.
Leishmaniasis is a tropical disease caused by an intracellular parasite of the genus Leishmania. The vector of transmission is the sand fly, which may deposit one of the 20 disease-causing protozoan species during blood ingestion. Clinical presentation depends on the complex interplay between the host cell-mediated immune response, and the specific protozoa and vector species. There are four generally accepted classifications of clinical disease: cutaneous, diffuse cutaneous, mucocutaneous and visceral leishmaniasis . According to the WHO, in 2013 about 215 thousand cases of leishmaniasis were reported worldwide.
Among the immunosensors reported, Sousa et al.  developed a new fluorescence-based immunosensor that comprised magnetic polymer microspheres coated with recombinant antigens, to improve the detection of anti-Leishmaniainfantum specific antibodies in the serum of infected dogs. Souto et al.  described the development of an immunosensor for anti-L. infantum antibodies based on detection by SPR technique.
Other kinds of transducers have been used, such as in the piezoelectric immunosensor developed for the detection of L. infantum antigens in tissues of infected hosts . In this case, antibodies were immobilized on a gold surface, covered with a thin film of cysteamine and glutaraldehyde, blocked with glycine and placed into contact with extracts of hamster spleens infected with L. infantum. The assay was able to detect 1.8 x 10-4amastigotes/g of infected tissue. Moreover, Mohan et al.  described an electrochemical genosensor based on the immobilization of a DNA sequence that targeted 18S rRNA gene sequences from Leishmaniadonovani.
Chagas disease, also known as American trypanosomiasis, is a neglected tropical disease caused by the protozoan parasite Trypanosomacruzi. It is found mainly in endemic areas of 21 Latin American countries, where it is mostly vector-borne transmitted to humans by contact with feces of haematophageous bugs of the Triatominae subfamily, known as ‘kissing bugs’, among other names, depending on the geographical area .
A polypeptide chain formed by recombinant antigens, cytoplasmic repetitive antigen and flagellar repetitive antigen of T. cruzi was adsorbed on gold and platinum electrodes and antigenantibody interaction was investigated by electrochemical impedance spectroscopy . T. cruzi proteins from epimastigote membranes were used for the construction of an amperometric immunosensor for serological diagnosis of Chagas disease. Antibodies present in the serum of patients were captured by the immobilized antigens and the affinity interaction was monitored by chronoamperometry using peroxidase-labeled IgG conjugate .
Ferreira et al.  described an amperometric bioelectrode for detection of antibodies occurring in sera of patients suffering from American trypanosomiasis. The strategy consisted of the immobilization of parasites proteins on goldelectrodes modified with thiol and the detection limit was 12.4 ng mL-1 of IgG. Pereira et al.  reported the development of an integrated microfluidic system coupled to a screen-printed carbon electrode applied to the quantitative determination of IgG specific antibodies present in serum samples. The electrode was modified by electrode position of gold nanoparticles and functionalized with T. cruzi proteins from epimastigote membranes. The calculated detection limit for electrochemical detection was 3.065 ng mL-1.
Toxoplasmosis is caused by the parasite Toxoplasma gondii, an obligate intracellular protozoan, capable of infecting humans. Most infections are asymptomatic or take a mild form, characterized by fever, malaise and lymphadenopathy. However, in cases of immune deficiency or when the parasite is congenitally acquired, it may cause serious illness and even death . The diagnosis and genetic characterization of T. gondii infection is crucial for monitoring, prevention and control of toxoplasmosis. Traditional approaches for the diagnosis of toxoplasmosis include molecular and imaging techniques .
Most biosensors for toxoplasmosis described in the literature are based on immunoassays for the detection of anti-T. gondii antibodies. An agglutination-based piezoelectric immunoassay was developed for directly detecting anti-T. gondii immunoglobulin’s in infected rabbit serum and blood. The proposed technique is based on the specific agglutination of antigen-coated gold nanoparticles (10 nm diameter), in the presence of the corresponding antibody, which causes a frequency change monitored by a piezoelectric device. The developed system is sensitive to dilution ratios of anti-T. gondii antibody as low as 1:5500 .
Ding et al.  developed an electrochemical biosensor based on an enzyme-catalyzed amplification. T. gondii antigen was immobilized on the surface of a gold electrode in order to bind anti-toxoplasma IgG, and this was followed by the addition of anti-toxoplasma IgG horseradish peroxidase conjugate. The transduction methods were quartz crystal microbalance, electrochemical impedance spectroscopy and cyclic voltammetry, with a detection limit of 1:9600 in dilution ratio.
Luo et al.  used two aptamers with high affinities to antitoxoplasma IgG as detection probes while developing a quantum dots-labeled dual aptasensor. In the presence of anti-toxoplasma IgG, an aptamer-protein-aptamer sandwich complex is formed and captured on a multi well microplate, whose fluorescence can be read out using quantum dots as label. The aptasensor has linearity within the range of 0.5-500 IU with the lowest detection of 0.1 IU. Also based on fluorimetric detection, He et al.  described the use of magnetic fluorescent nanoparticles in the development of a genosensor for the detection of T. gondii DNA oligonucleotides, with limit of detection of 8.339 nM.
Since the first ideas five decades ago, biosensors have shown their potential to revolutionize the diagnosis of a variety of health conditions. Today, their impact in clinical management is well established, since rapidity, specificity and sensibility are crucial characteristics for early diagnosis and therapy initiation. The development of new technologies, such as nanotechnology and microfluidics, together with biomarker discovery should improve their effectiveness. In the case of infectious diseases, which have the potential of transmission and outbreaks occurrence, with possible sequels development and lethality, the availability of robust diagnosis methods is crucial.
Biosensors are also important in the democratization of diagnosis. Many methods currently available are inaccessible for a significant part of the world population, since they are expensive, centralized and require specialized technicians for operation. Therefore, the potential of cost reduction, portability and simplicity is largely appreciable, especially in the case of neglected diseases.
What lies ahead is also promising. Advances in fields such as genetics and epigenetics, chemistry and biochemistry, physiology and bioinformatics have the potential of clarifying the nuances of biological processes in the health and disease. New targets of study are emerging and other are being better understood, especially in diagnosis and therapeutics, or even both (i.e., theranostics). Therefore, the coupling of these findings with promising technologies such as biosensors may change the current landscape of clinical diagnosis.
- Neiderud CJ. How urbanization affects the epidemiology of emerging infectious diseases. Infect Ecol Epidemiol. 2015; 5: 27060.
- Van Panhuis WG, Grefenstette J, Jung SY, Chok NS, Cross A, Eng H, et al. Contagious diseases in the United States from 1888 to the present. N Engl J Med. 2013; 369: 2152-2158.
- Dye C. After 2015: infectious diseases in a new era of health and development. Philos Trans R Soc Lond B Biol Sci. 2014; 369: 20130426.
- Almeida SL. Trending now: re-emerging infectious disease update. J Emerg Nurs. 2015; 41: 104-108.
- TDR Diagnostics Evaluation Expert Panel. Banoo S, Bell D, Bossuyt P, Herring A, Mabey D, Poole F, et al. Evaluation of diagnostic tests for infectious diseases: general principles. Nat Rev Microbiol. 2010; 8: 17-29.
- Carinelli S, MartÃ M, Alegret S, Pividori MI. Biomarker detection of global infectious diseases based on magnetic particles. N Biotechnol. 2015; 32: 521-532.
- Ince J, McNally A. Development of rapid, automated diagnostics for infectious disease: advances and challenges. Expert Rev Med Devices. 2009; 6: 641-651.
- Ko ER, Yang WE, McClain MT, Woods CW, Ginsburg GS, Tsalik EL. What was old is new again: using the host response to diagnose infectious disease. Expert Rev Mol Diagn. 2015; 15: 1143-1158.
- Heffernan DS, Fox ED. Advancing technologies for the diagnosis and management of infections. Surg Clin North Am. 2014; 94: 1163-1174.
- Patel R. MALDI-TOF MS for the diagnosis of infectious diseases. Clin Chem. 2015; 61: 100-111.
- Calderaro A, Arcangeletti MC, Rodighiero I, Buttrini M, Gorrini C, Motta F, et al. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry applied to virus identification. Sci Rep. 2014; 4: 6803.
- Thorburn F, Bennett S, Modha S, Murdoch D, Gunson R, Murcia PR. The use of next generation sequencing in the diagnosis and typing of respiratory infections. J Clin Virol. 2015; 69: 96-100.
- Barzon L, Lavezzo E, Costanzi G, Franchin E, Toppo S, PalÃ¹ G. Next-generation sequencing technologies in diagnostic virology. J Clin Virol. 2013; 58: 346-350.
- Su W, Gao X, Jiang L, Qin J. Microfluidic platform towards point-of-care diagnostics in infectious diseases. J Chromatogr A. 2015; 1377: 13-26.
- Qasim M, Lim DJ, Park H, Na D. Nanotechnology for diagnosis and treatment of infectious diseases. J Nanosci Nanotechnol. 2014; 14: 7374-7387.
- Biomarkers Definitions Working Group. Biomarkers and surrogate endpoints: preferred definitions and conceptual framework. Clin Pharmacol Ther. 2001; 69: 89-95.
- Wehrens R, Franceschi P, Vrhovsek U, Mattivi F. Stability-based biomarker selection. Anal Chim Acta. 2011; 705: 15-23.
- Naylor S. Biomarkers: current perspectives and future prospects. Expert Rev Mol Diagn. 2003; 3: 525-529.
- Gupta S, Venkatesh A, Ray S, Srivastava S9*9. Challenges and prospects for biomarker research: a current perspective from the developing world. Biochim Biophys Acta. 2014; 1844: 899-908.
- He Z, Yu W. Stable feature selection for biomarker discovery. Comput Biol Chem. 2010; 34: 215-225.
- Goulart LR, Vieira CU, Freschi AP, Capparelli FE, Fujimura PT, Almeida JF, et al. Biomarkers for serum diagnosis of infectious diseases and their potential application in novel sensor platforms. Crit Rev Immunol. 2010; 30: 201-222.
- Loonen AJ, Wolffs PF, Bruggeman CA, van den Brule AJ. Developments for improved diagnosis of bacterial bloodstream infections. Eur J Clin Microbiol Infect Dis. 2014; 33: 1687-1702.
- Scheler O, Glynn B, Kurg A. Nucleic acid detection technologies and marker molecules in bacterial diagnostics. Expert Rev Mol Diagn. 2014; 14: 489-500.
- Chuang LY, Cheng YH, Yang CH. Specific primer design for the polymerase chain reaction. Biotechnol Lett. 2013; 35: 1541-1549.
- Noguera DR, Wright ES, Camejo P, Yilmaz LS. Mathematical tools to optimize the design of oligonucleotide probes and primers. Appl Microbiol Biotechnol. 2014; 98: 9595-9608.
- Guo J, Ju J, Turro NJ. Fluorescent hybridization probes for nucleic acid detection. Anal Bioanal Chem. 2012; 402: 3115-3125.
- Xia XQ, Jia Z, Porwollik S, Long F, Hoemme C, Ye K, et al. Evaluating oligonucleotide properties for DNA microarray probe design. Nucleic Acids Res. 2010; 38: 121.
- Cho MS, Ahn TY, Joh K, Paik SY, Kwon OS, Jheong WH, et al. A novel marker for the species-specific detection and quantitation of Vibrio cholerae by targeting an outer membrane lipoprotein lolB gene. J Microbiol Biotechnol. 2013; 23: 555-559.
- Ogura A, Yoshida MA, Fukuzaki M, Sese J. In vitro homology search array comprehensively reveals highly conserved genes and their functional characteristics in non-sequenced species. BMC Genomics. 2010; 11: 9.
- Naidoo N, Ramsugit S, Pillay M. Mycobacterium tuberculosis pili (MTP), a putative biomarker for a tuberculosis diagnostic test. Tuberculosis (Edinb). 2014; 94: 338-345.
- Natesan M, Ulrich RG. Protein microarrays and biomarkers of infectious disease. Int J Mol Sci. 2010; 11: 5165-5183.
- Carmona SJ, Sartor PA, Leguizamon MS, Campetella OE, Aguero F. Diagnostic peptide discovery: prioritization of pathogen diagnostic markers using multiple features. PLoS One. 2012; 7: 50748.
- Pande J, Szewczyk MM, Grover AK. Phage display: concept, innovations, applications and future. Biotechnol Adv. 2010; 28: 849-858.
- Alban SM, de Moura JF, Thomaz-Soccol V, Buhrer Sekula S, Alvarenga LM, Mira MT, et al. Phage display and synthetic peptides as promising biotechnological tools for the serological diagnosis of leprosy. PLoS One. 2014; 9: 106222.
- Costa LE, Goulart LR, Pereira NC, Lima MI, Duarte MC, Martins VT, et al. Mimotope-based vaccines of Leishmania infantum antigens and their protective efficacy against visceral leishmaniasis. PLoS One. 2014; 9: 110014.
- Wu J, Zeng XQ, Zhang HB, Ni HZ, Pei L, Zou LR, et al. Novel phage display-derived H5N1-specific scFvs with potential use in rapid avian flu diagnosis. J Microbiol Biotechnol. 2014; 24: 704-713.
- Lim BN, Tye GJ, Choong YS, Ong EB, Ismail A, Lim TS. Principles and application of antibody libraries for infectious diseases. Biotechnol Lett. 2014; 36: 2381-2392.
- Shimoni M, Herschhorn A, Britan-Rosich Y, Kotler M, Benhar I, Hizi A. The isolation of novel phage display-derived human recombinant antibodies against CCR5, the major co-receptor of HIV. Viral Immunol. 2013; 26: 277-290.
- Zimbres FM, Tárnok A, Ulrich H, Wrenger C. Aptamers: novel molecules as diagnostic markers in bacterial and viral infections? Biomed Res Int. 2013; 731516.
- Xi Z, Huang R, Deng Y, He N. Progress in selection and biomedical applications of aptamers. J Biomed Nanotechnol. 2014; 10: 3043-3062.
- Aimaiti R, Qin L, Cao T, Yang H, Wang J, Lu J, et al. Identification and application of ssDNA aptamers against HÃ¢â€šÆ’Ã¢â€šâ€¡Rv in the detection of Mycobacterium tuberculosis. Appl Microbiol Biotechnol. 2015; 99: 9073-9083.
- Shiratori I, Akitomi J, Boltz DA, Horii K, Furuichi M, Waga I. Selection of DNA aptamers that bind to influenza A viruses with high affinity and broad subtype specificity. Biochem Biophys Res Commun. 2014; 443: 37-41.
- Chen HL, Hsiao WH, Lee HC, Wu SC, Cheng JW. Selection and Characterization of DNA Aptamers Targeting All Four Serotypes of Dengue Viruses. PLoS One. 2015; 10: e0131240.
- Jain P, Chakma B, Patra S, Goswami P. Potential biomarkers and their applications for rapid and reliable detection of malaria. Biomed Res Int. 2014; 2014: 852645.
- Wang J. Electrochemical biosensors: towards point-of-care cancer diagnostics. Biosens Bioelectron. 2006; 21: 1887-1892.
- Vopálenská I, Váchová L, Palková Z. New biosensor for detection of copper ions in water based on immobilized genetically modified yeast cells. Biosens Bioelectron. 2015; 72: 160-167.
- Hughes G, Pemberton RM, Fielden PR, Hart JP. Development of a novel reagentless, screen-printed amperometric biosensor based on glutamate dehydrogenase and NAD+, integrated with multi-walled carbon nanotubes for the determination of glutamate in food and clinical applications. Sens Actuators B Chem. 2015; 216: 614-621.
- Kashem MA, Suzuki M, Kimoto K, Iribe Y. An optical biochemical oxygen demand biosensor chip for environmental monitoring. Sens Actuators B Chem. 2015.
- Nasirizadeh N, Zare HR, Pournaghi-Azar MH, Hejazi MS. Introduction of hematoxylin as an electroactive label for DNA biosensors and its employment in detection of target DNA sequence and single-base mismatch in human papilloma virus corresponding to oligonucleotide. Biosens Bioelectron. 2011; 26: 2638-2644.
- Balvedi RP, Castro AC, Madurro JM, Brito-Madurro AG. Detection of a specific biomarker for Epstein-Barr virus using a polymer-based genosensor. Int J Mol Sci. 2014; 15: 9051-9066.
- Nambiar S, Yeow JT. Conductive polymer-based sensors for biomedical applications. Biosens Bioelectron. 2011; 26: 1825-1832.
- Tymecki L, Koncki R. Thick-film potentiometric biosensor for bloodless monitoring of hemodialysis. Sens Actuators B Chem. 2006; 113: 782-786.
- Chen JC, Chung HH, Hsu CT, Tsai DM, Kumar AS, Zen JM. A disposable single-use electrochemical sensor for the detection of uric acid in human whole blood. Sens Actuators B Chem. 2005; 110: 364-369.
- Zhang Y, Tadigadapa S. Calorimetric biosensors with integrated microfluidic channels. Biosens Bioelectron. 2004; 19: 1733-1743.
- Su L, Zou L, Fong CC, Wong WL, Wei F, Wong KY, et al. Detection of cancer biomarkers by piezoelectric biosensor using PZT ceramic resonator as the transducer. Biosens Bioelectron. 2013; 46: 155-161.
- Pahurkar VG, Tamgadge YS, Gambhire AB, Muley GG. Glucose oxidase immobilized PANI cladding modified fiber optic intrinsic biosensor for detection of glucose. Sens Actuators B Chem. 2015; 210: 362-368.
- Gerard M, Chaubey A, Malhotra BD. Application of conducting polymers to biosensors. Biosens Bioelectron. 2002; 17: 345-359.
- Sheng LP, Lertanantawong B, Yin LS, Ravichandran M, Heng LY, Surareungchai W. Electrochemical genosensor assay using lyophilized gold nanoparticles/latex microsphere label for detection of Vibrio cholerae. Talanta. 2015; 139: 167-173.
- Shen Y, Zhang Y, Liu M, Liu X, Guo H, Zhang X, et al. A simple and sensitive electrochemical immunosensor based on thiol aromatic aldehyde as a substrate for the antibody immobilization. Talanta. 2015; 141: 288-292.
- Clark LC, Lyons C. Electrode systems for continuous monitoring in cardiovascular surgery. Ann N Y Acad Sci. 1962; 102: 29-45.
- Murphy L. Biosensors and bioelectrochemistry. Curr Opin Chem Biol. 2006; 10: 177-184.
- Castro ACH, FranÃ§a EG, de Paula LF, Soares MMCN, Goulart LR, Madurro JM, et al. Preparation of genosensor for detection of specific DNA sequence of the hepatitis B virus. Applied Surface Science. 2014; 314: 273-279.
- Ligaj M, Tichoniuk M, Gwiazdowska D, Filipiak M. Electrochemical DNA biosensor for the detection of pathogenic bacteria Aeromonas hydrophila. Electrochimica Acta. 2014; 128: 67-74.
- Luz JGG, Souto DEP, Machado-Assis GF, de Lana M, Kubota LT, Luz RCS, et al. Development and evaluation of a SPR-based immunosensor for detection of anti-Trypanosoma cruzi antibodies in human serum. Sens Actuators B Chem. 2015; 212: 287-296.
- Dmitriev DA, Massino YS, Segal OL, Smirnova MB, Pavlova EV, Gurevich KG, et al. Analysis of the binding of bispecific monoclonal antibodies with immobilized antigens (human IgG and horseradish peroxidase) using a resonant mirror biosensor. J Immunol Methods. 2002; 261: 103-118.
- Abe K, Yoshida W, Ikebukuro K. Electrochemical biosensors using aptamers for theranostics. Adv Biochem Eng Biotechnol. 2014; 140: 183-202.
- Sanders CA, Rodriguez M, Greenbaum E. Stand-off tissue-based biosensors for the detection of chemical warfare agents using photosynthetic fluorescence induction. Biosens Bioelectron. 2001; 16: 439-446.
- Park M, Tsai SL, Chen W. Microbial biosensors: engineered microorganisms as the sensing machinery. Sensors (Basel). 2013; 13: 5777-5795.
- Ensafi AA, Taei M, Rahmani HR, Khayamian T. Sensitive DNA impedance biosensor for detection of cancer, chronic lymphocytic leukemia, based on gold nanoparticles/gold modified electrode. Electrochimica Acta. 2011; 56: 8176-8183.
- Ensafi AA, Amini M, Rezaei B. Detection of DNA damage induced by chromium/glutathione/H2O2 system at MWCNTsâ€“poly(diallyldimethylammonium chloride) modified pencil graphite electrode using methylene blue as an electroactive probe. Sensors and Actuators B: Chemical. 2013; 177: 862-870.
- Ensafi AA, Amini M, Rezaei B, Talebi M. A novel diagnostic biosensor for distinguishing immunoglobulin mutated and unmutated types of chronic lymphocytic leukemia. Biosens Bioelectron. 2016; 77: 409-415.
- Ensafi AA, Amini M, Rezaei B. Impedimetric DNA-biosensor for the study of anti-cancer action of mitomycin C: comparison between acid and electroreductive activation. Biosens Bioelectron. 2014; 59: 282-288.
- Kim K, Park C, Kwon D, Kim D, Meyyappan M, Jeon S, et al. Silicon nanowire biosensors for detection of cardiac troponin I (cTnI) with high sensitivity. Biosens Bioelectron. 2016; 77: 695-701.
- Kucherenko IS, Soldatkin OO, Lagarde F, Jaffrezic-Renault N, Dzyadevych SV, Soldatkin AP. Determination of total creatine kinase activity in blood serum using an amperometric biosensor based on glucose oxidase and hexokinase. Talanta. 2015; 144: 604-611.
- Lee HY, Choi JS, Guruprasath P, Lee BH, Cho YW. An Electrochemical Biosensor Based on a Myoglobin-specific Binding Peptide for Early Diagnosis of Acute Myocardial Infarction. Analytical Sciences. 2015; 31: 699-704.
- Beleoken E, Leh H, Arnoux A, Ducot B, Nogues C, De Martin E, et al. SPRi-based strategy to identify specific biomarkers in systemic lupus erythematosus, rheumatoid arthritis and autoimmune hepatitis. PLoS One. 2013; 8: 84600.
- Park YM, Kim SJ, Lee KJ, Yang SS, Min BH, Yoon HC. Detection of CTX-II in serum and urine to diagnose osteoarthritis by using a fluoro-microbeads guiding chip. Biosens Bioelectron. 2015; 67: 192-199.
- Villa Mde G, Jimenez-Jorquera C, Haro I, Gomara MJ, Sanmarti R, Fernandez-Sanchez C, et al. Carbon nanotube composite peptide-based biosensors as putative diagnostic tools for rheumatoid arthritis. Biosens Bioelectron. 2011; 27: 113-118.
- Babu E, Muthu Mareeswaran P, Sathish V, Singaravadivel S, Rajagopal S. Sensing and inhibition of amyloid-îÂ² based on the simple luminescent aptamer-ruthenium complex system. Talanta. 2015; 134: 348-353.
- Esteves-Villanueva JO, Trzeciakiewicz H, Martic S. A protein-based electrochemical biosensor for detection of tau protein, a neurodegenerative disease biomarker. Analyst. 2014; 139: 2823-2831.
- Kim JA, Kim M, Kang SM, Lim KT, Kim TS, Kang JY. Magnetic bead droplet immunoassay of oligomer amyloidîÂ² for the diagnosis of Alzheimer's disease using micro-pillars to enhance the stability of the oil-water interface. Biosens Bioelectron. 2015; 67: 724-732.
- Zhao Z, Zhu L, Bu X, Ma H, Yang S, Yang Y, et al. Label-free detection of Alzheimer's disease through the ADP3 peptoid recognizing the serum amyloid-beta42 peptide. Chem Commun (Camb). 2015; 51: 718-721.
- Huang H, Bai W, Dong C, Guo R, Liu Z. An ultrasensitive electrochemical DNA biosensor based on graphene/Au nanorod/polythionine for human papillomavirus DNA detection. Biosens Bioelectron. 2015; 68: 442-446.
- Cecchetto J, Carvalho FC, Santos A, Fernandes FCB, Bueno PR. An impedimetric biosensor to test neat serum for dengue diagnosis. Sens Actuators B Chem. 2015; 213: 150-154.
- Nguyen BT, Peh AE, Chee CY, Fink K, Chow VT, Ng MM, et al. Electrochemical impedance spectroscopy characterization of nanoporous alumina dengue virus biosensor. Bioelectrochemistry. 2012; 88: 15-21.
- Zhang B, Salieb-Beugelaar GB, Nigo MM, Weidmann M, Hunziker P. Diagnosing dengue virus infection: rapid tests and the role of micro/nanotechnologies. Nanomedicine. 2015; 11: 1745-1761.
- Choi YH, Lee GY, Ko H, Chang YW, Kang MJ, Pyun JC. Development of SPR biosensor for the detection of human hepatitis B virus using plasma-treated parylene-N film. Biosens Bioelectron. 2014; 56: 286-294.
- Shourian M, Ghourchian H, Boutorabi M. Ultra-sensitive immunosensor for detection of hepatitis B surface antigen using multi-functionalized gold nanoparticles. Anal Chim Acta. 2015; 895: 1-11.
- Yang L, Du F, Chen G, Yasmeen A, Tang Z. A novel colorimetric PCR-based biosensor for detection and quantification of hepatitis B virus. Anal Chim Acta. 2014; 840: 75-81.
- Campos-Ferreira DS, Nascimento GA, Souza EV, Souto-Maior MA, Arruda MS, Zanforlin DM, et al. Electrochemical DNA biosensor for human papillomavirus 16 detection in real samples. Anal Chim Acta. 2013; 804: 258-263.
- Urrego LF, Lopez DI, Ramirez KA, Ramirez C, Osma JF. Biomicrosystem design and fabrication for the human papilloma virus 16 detection. Sens Actuators B Chem. 2015; 207: 97-104.
- Elzouki AN, Elgamay SM, Zorgani A, Elahmer O. Hepatitis B and C status among health care workers in the five main hospitals in eastern Libya. J Infect Public Health. 2014; 7: 534-541.
- Zhang C, Lou J, Tu W, Bao J, Dai Z. Ultrasensitive electrochemical biosensing for DNA using quantum dots combined with restriction endonuclease. Analyst. 2015; 140: 506-511.
- Nascimento HP, Oliveira MD, de Melo CP, Silva GJ, Cordeiro MT, Andrade CA. An impedimetric biosensor for detection of dengue serotype at picomolar concentration based on gold nanoparticles-polyaniline hybrid composites. Colloids Surf B Biointerfaces. 2011; 86: 414-419.
- Silva MM, Dias AC, Cordeiro MT, Marques E, Goulart MO, Dutra RF. A thiophene-modified screen printed electrode for detection of dengue virus NS1 protein. Talanta. 2014; 128: 505-510.
- Sanvicens N, Pastells C, Pascual N, Marco MP. Nanoparticle-based biosensors for detection of pathogenic bacteria. TrAC Trends in Analytical Chemistry. 2009; 28: 1243-1252.
- Wang Y, Ye Z, Ying Y. New trends in impedimetric biosensors for the detection of foodborne pathogenic bacteria. Sensors (Basel). 2012; 12: 3449-3471.
- Singh R, Mukherjee MD, Sumana G, Gupta RK, Sood S, Malhotra BD. Biosensors for pathogen detection: A smart approach towards clinical diagnosis. Sens Actuators B Chem. 2014; 197: 385-404.
- Singh A, Poshtiban S, Evoy S. Recent advances in bacteriophage based biosensors for food-borne pathogen detection. Sensors (Basel). 2013; 13: 1763-1786.
- Ibrahim F, Thio TH, Faisal T. The application of biomedical engineering techniques to the diagnosis and management of tropical diseases: a review. Sensors (Basel). 2015; 15: 6947-6995.
- Garcia-Monco JC. Tuberculosis. Handb Clin Neurol. 2014; 121: 1485-1499.
- Liu C, Jiang D, Xiang G, Liu L, Liu F, Pu X. An electrochemical DNA biosensor for the detection of Mycobacterium tuberculosis, based on signal amplification of graphene and a gold nanoparticle-polyaniline nanocomposite. Analyst. 2014; 139: 5460-5465.
- Costa MP, Andrade CA, Montenegro RA, Melo FL, Oliveira MD. Self-assembled monolayers of mercaptobenzoic acid and magnetite nanoparticles as an efficient support for development of tuberculosis genosensor. J Colloid Interface Sci. 2014; 433: 141-148.
- Barreda-Garcia S, Gonzalez-Alvarez MJ, de-los-Santos-Alvarez N, Palacios-Gutierrez JJ, Miranda-Ordieres AJ, Lobo-Castanon MJ. Attomolar quantitation of Mycobacterium tuberculosis by asymmetric helicase-dependent isothermal DNA-amplification and electrochemical detection. Biosens Bioelectron. 2015; 68: 122-128.
- Rachkov A, Patskovsky S, Soldatkin A, Meunier M. Discrimination of single base mismatched oligonucleotides related to the rpoB gene of Mycobacterium tuberculosis using a surface plasmon resonance biosensor. Biotechnol Appl Biochem. 2013; 60: 453-458.
- Hsu SH, Lin YY, Lu SH, Tsai IF, Lu YT, Ho HT. Mycobacterium tuberculosis DNA detection using surface plasmon resonance modulated by telecommunication wavelength. Sensors (Basel). 2013; 14: 458-467.
- Xiang Y, Zhu X, Huang Q, Zheng J, Fu W. Real-time monitoring of mycobacterium genomic DNA with target-primed rolling circle amplification by a Au nanoparticle-embedded SPR biosensor. Biosens Bioelectron. 2015; 66: 512-519.
- Hsieh SC, Chang CC, Lu CC, Wei CF, Lin CS, Lai HC, et al. Rapid identification of Mycobacterium tuberculosis infection by a new array format-based surface plasmon resonance method. Nanoscale Res Lett. 2012; 7: 180.
- Mukundan H, Kumar S, Price DN, Ray SM, Lee YJ, Min S, et al. Rapid detection of Mycobacterium tuberculosis biomarkers in a sandwich immunoassay format using a waveguide-based optical biosensor. Tuberculosis (Edinb). 2012; 92: 407-416.
- Kim JH, Yeo WH, Shu Z, Soelberg SD, Inoue S, Kalyanasundaram D, et al. Immunosensor towards low-cost, rapid diagnosis of tuberculosis. Lab Chip. 2012; 12: 1437-1440.
- Hiraiwa M, Kim JH, Lee HB, Inoue S, Becker AL, Weigel KM, et al. Amperometric immunosensor for rapid detection of Mycobacterium tuberculosis. J Micromech Microeng. 2015; 25: 055013.
- Goulart IM, Goulart LR. Leprosy: diagnostic and control challenges for a worldwide disease. Arch Dermatol Res. 2008; 300: 269-290.
- Afonso AS, Goulart LR, Goulart IMB, Machado AEH, Madurro JM, Brito-Madurro AG. A promising bioelectrode based on gene of Mycobacterium leprae immobilized onto poly(4-aminophenol). J Appl Polym Sci. 2010; 118: 2921-2928.
- Paula Vaz Cardoso L, Dias RF, Freitas AA, Hungria EM, Oliveira RM, Collovati M, et al. Development of a quantitative rapid diagnostic test for multibacillary leprosy using smart phone technology. BMC Infect Dis. 2013; 13: 497.
- Abio A, Neal KR, Beck CR. An epidemiological review of changes in meningococcal biology during the last 100 years. Pathog Glob Health. 2013; 107: 373-380.
- Bahr NC, Boulware DR. Methods of rapid diagnosis for the etiology of meningitis in adults. Biomark Med. 2014; 8: 1085-1103.
- Reddy SB, Mainwaring DE, Kobaisi MA, Zeephongsekul P, Fecondo JV. Acoustic wave immunosensing of a meningococcal antigen using gold nanoparticle-enhanced mass sensitivity. Biosens Bioelectron. 2012; 31: 382-387.
- Dash SK, Sharma M, Khare S, Kumar A. Omp85 genosensor for detection of human brain bacterial meningitis. Biotechnol Lett. 2013; 35: 929-935.
- Patel MK, Solanki PR, Seth S, Gupta S, Khare S, Kumar A, et al. CtrA gene based electrochemical DNA sensor for detection of meningitis. Electrochem Commun. 2009; 11: 969-973.
- Patel MK, Solanki PR, Kumar A, Khare S, Gupta S, Malhotra BD. Electrochemical DNA sensor for Neisseria meningitidis detection. Biosens Bioelectron. 2010; 25: 2586-2591.
- Tak M, Gupta V, Tomar M. Flower-like ZnO nanostructure based electrochemical DNA biosensor for bacterial meningitis detection. Biosens Bioelectron. 2014; 59: 200-207.
- Dash SK, Sharma M, Khare S, Kumar A. rmpM genosensor for detection of human brain bacterial meningitis in cerebrospinal fluid. Appl Biochem Biotechnol. 2013; 171: 198-208.
- Sharma MK, Agarwal GS, Rao VK, Upadhyay S, Merwyn S, Gopalan N, et al. Amperometric immunosensor based on gold nanoparticles/alumina sol-gel modified screen-printed electrodes for antibodies to Plasmodium falciparum histidine rich protein-2. Analyst. 2010; 135: 608-614.
- Sharma MK, Rao VK, Merwyn S, Agarwal GS, Upadhyay S, Vijayaraghavan R. A novel piezoelectric immunosensor for the detection of malarial Plasmodium falciparum histidine rich protein-2 antigen. Talanta. 2011; 85: 1812-1817.
- Sharma MK, Rao VK, Agarwal GS, Rai GP, Gopalan N, Prakash S, et al. Highly sensitive amperometric immunosensor for detection of Plasmodium falciparum histidine-rich protein 2 in serum of humans with malaria: comparison with a commercial kit. J Clin Microbiol. 2008; 46: 3759-3765.
- Sikarwar B, Sharma PK, Srivastava A, Agarwal GS, Boopathi M, Singh B, et al. Surface plasmon resonance characterization of monoclonal and polyclonal antibodies of malaria for biosensor applications. Biosens Bioelectron. 2014; 60: 201-209.
- Lee S, Song KM, Jeon W, Jo H, Shim YB, Ban C. A highly sensitive aptasensor towards Plasmodium lactate dehydrogenase for the diagnosis of malaria. Biosens Bioelectron. 2012; 35: 291-296.
- Jeon W, Lee S, Manjunatha DH, Ban C. A colorimetric aptasensor for the diagnosis of malaria based on cationic polymers and gold nanoparticles. Anal Biochem. 2013; 439: 11-16.
- Lee S, Manjunatha DH, Jeon W, Ban C. Cationic surfactant-based colorimetric detection of Plasmodium lactate dehydrogenase, a biomarker for malaria, using the specific DNA aptamer. PLoS One. 2014; 9: 100847.
- Ittarat W, Chomean S, Sanchomphu C, Wangmaung N, Promptmas C, Ngrenngarmlert W. Biosensor as a molecular malaria differential diagnosis. Clin Chim Acta. 2013; 419: 47-51.
- Kevric I, Cappel MA, Keeling JH. New World and Old World Leishmania Infections: A Practical Review. Dermatol Clin. 2015; 33: 579-593.
- Sousa S, Cardoso L, Reed SG, Reis AB, Martins-Filho OA, Silvestre R, et al. Development of a fluorescent based immunosensor for the serodiagnosis of canine leishmaniasis combining immunomagnetic separation and flow cytometry. PLoS Negl Trop Dis. 2013; 7: 2371.
- Souto DE, Silva JV, Martins HR, Reis AB, Luz RC, Kubota LT, et al. Development of a label-free immunosensor based on surface plasmon resonance technique for the detection of anti-Leishmania infantum antibodies in canine serum. Biosens Bioelectron. 2013; 46: 22-29.
- Cabral-Miranda G, de Jesus JR, Oliveira PR, Britto GS, Pontes-de-Carvalho LC, Dutra RF, et al. Detection of parasite antigens in Leishmania infantum-infected spleen tissue by monoclonal antibody-, piezoelectric-based immunosensors. J Parasitol. 2014; 100: 73-78.
- Mohan S, Srivastava P, Maheshwari SN, Sundar S, Prakash R. Nano-structured nickel oxide based DNA biosensor for detection of visceral leishmaniasis (Kala-azar). Analyst. 2011; 136: 2845-2851.
- Nouvellet P, CucunubÃ¡ ZM, GourbiÃ¨re S. Ecology, evolution and control of Chagas disease: a century of neglected modelling and a promising future. Adv Parasitol. 2015; 87: 135-191.
- Diniz FB, Ueta RR, Pedrosa AM, Areias Mda C, Pereira VR, Silva ED, et al. Impedimetric evaluation for diagnosis of Chagas' disease: antigen-antibody interactions on metallic electrodes. Biosens Bioelectron. 2003; 19: 79-84.
- Ferreira AA, Colli W, da Costa PI, Yamanaka H. Immunosensor for the diagnosis of Chagas' disease. Biosens Bioelectron. 2005; 21: 175-181.
- Pereira SV, Bertolino FA, Fernandez-Baldo MA, Messina GA, Salinas E, Sanz MI, et al. A microfluidic device based on a screen-printed carbon electrode with electrodeposited gold nanoparticles for the detection of IgG anti-Trypanosoma cruzi antibodies. Analyst. 2011; 136: 4745-4751.
- Xiao J, Yolken RH. Strain hypothesis of Toxoplasma gondii infection on the outcome of human diseases. Acta Physiol (Oxf). 2015; 213: 828-845.
- Wang H, Lei C, Li J, Wu Z, Shen G, Yu R. A piezoelectric immunoagglutination assay for Toxoplasma gondii antibodies using gold nanoparticles. Biosens Bioelectron. 2004; 19: 701-709.
- Ding Y, Wang H, Shen G, Yu R. Enzyme-catalyzed amplified immunoassay for the detection of Toxoplasma gondii-specific IgG using Faradaic impedance spectroscopy, CV and QCM. Anal Bioanal Chem. 2005; 382: 1491-1499.
- Luo Y, Liu X, Jiang T, Liao P, Fu W. Dual-aptamer-based biosensing of toxoplasma antibody. Anal Chem. 2013; 85: 8354-8360.
- He L, Ni L, Zhang X, Zhang C, Li R, Xu S. Fluorescent Detection of Specific DNA Sequences Related to Toxoplasma gondii Based on Magnetic Fluorescent Nanoparticles Fe3O4/CdTe Biosensor. Int J Biochem Res Rev. 2015; 6: 130-139.
- Ronkainen NJ, Halsall HB, Heineman WR. Electrochemical biosensors. Chem Soc Rev. 2010; 39: 1747-1763.
- Wohlfahrt G, Witt S, Hendle J, Schomburg D, Kalisz HM, Hecht HJ. 1.8 and 1.9 A resolution structures of the Penicillium amagasakiense and Aspergillus niger glucose oxidases as a basis for modelling substrate complexes. Acta Crystallogr D Biol Crystallogr. 1999; 55: 969-977.
- Saphire EO, Parren PW, Pantophlet R, Zwick MB, Morris GM, Rudd PM, et al. Crystal structure of a neutralizing human IGG against HIV-1: a template for vaccine design. Science. 2001; 293: 1155-1159.
- Drew HR, Wing RM, Takano T, Broka C, Tanaka S, Itakura K, et al. Structure of a B-DNA dodecamer: conformation and dynamics. Proc Natl Acad Sci U S A. 1981; 78: 2179-2183.
Citation: Rodovalho VR, Alves LM, Castro ACH, Madurro JM, Brito-Madurro AG and Santos AR. Biosensors Applied to Diagnosis of Infectious Diseases â€“ An Update. Austin J Biosens & Bioelectron. 2015; 1(3): 1015. ISSN : 2473-0629