An ELISA using a recombinant chimera of protective antigen and lethal factor for serodiagnosis of cutaneous anthrax in India

Varshney, A.; Puranik, Nidhi; Kumar, Manoj; Pal, Vijai; Padmaja, Jyothi; Goel, Ajay

doi:10.1016/j.biologicals.2019.01.001

Cited by 8 publications

(5 citation statements)

References 33 publications

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“…EF can convert up to 2000 molecules of ATP to cAMP per second, significantly increasing the concentration of this classical second messenger, thereby causing diverse effects [52]. There are three forms of anthrax (inhalational, gastrointestinal, and cutaneous) depending on the route of exposure to B. anthracis [53]. Inhalational anthrax is considered to be the most fatal form of anthrax.…”

Section: Bacillus Anthracismentioning

confidence: 99%

Dangerous Pathogens as a Potential Problem for Public Health

et al. 2020

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Pathogens are various organisms, such as viruses, bacteria, fungi, and protozoa, which can cause severe illnesses to their hosts. Throughout history, pathogens have accompanied human populations and caused various epidemics. One of the most significant outbreaks was the Black Death, which occurred in the 14th century and caused the death of one-third of Europe’s population. Pathogens have also been studied for their use as biological warfare agents by the former Soviet Union, Japan, and the USA. Among bacteria and viruses, there are high priority agents that have a significant impact on public health. Bacillus anthracis, Francisella tularensis, Yersinia pestis, Variola virus, Filoviruses (Ebola, Marburg), Arenoviruses (Lassa), and influenza viruses are included in this group of agents. Outbreaks and infections caused by them might result in social disruption and panic, which is why special operations are needed for public health preparedness. Antibiotic-resistant bacteria that significantly impede treatment and recovery of patients are also valid threats. Furthermore, recent events related to the massive spread of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are an example of how virus-induced diseases cannot be ignored. The impact of outbreaks, such as SARS-CoV-2, have had far-reaching consequences beyond public health. The economic losses due to lockdowns are difficult to estimate, but it would take years to restore countries to pre-outbreak status. For countries affected by the 2019 coronavirus disease (COVID-19), their health systems have been overwhelmed, resulting in an increase in the mortality rate caused by diseases or injuries. Furthermore, outbreaks, such as SARS-CoV-2, will induce serious, wide-ranging (and possibly long-lasting) psychological problems among, not only health workers, but ordinary citizens (this is due to isolation, quarantine, etc.). The aim of this paper is to present the most dangerous pathogens, as well as general characterizations, mechanisms of action, and treatments.

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Section: Bacillus Anthracismentioning

confidence: 99%

Dangerous Pathogens as a Potential Problem for Public Health

et al. 2020

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“…Laboratory testing is required to confirm the diagnosis, and the detection of anthrax protective antigen (PA) has been found to be an important biological marker for anthrax [5]. However, existing PA detection technologies include enzyme-linked immunosorbent assays (ELISAs) [6], immunomagnetic beads (IMBs) [7,8], homogenous time-resolved fluorescence (HTRF) [9,10], and biosensors [11][12][13][14], which can only detect nanogram-or picogram-level concentrations of PA and have problems such as cumbersome operation steps, a large instrument volume, and high consumable prices that cannot be solved in the short term.…”

Section: Introductionmentioning

confidence: 99%

Research on Detection of Ultra-Low Concentration Anthrax Protective Antigen Using Graphene Field-Effect Transistor Biosensor

Liang¹,

Chen²,

Yan³

et al. 2023

Sensors

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Background: Protective antigen (PA) is an important biomarker for the early diagnosis of anthrax, and the accurate detection of protective antigen under extremely low concentration conditions has always been a hot topic in the biomedical field. To complete the diagnosis of anthrax in a timely manner, it is necessary to detect PA at extremely low concentrations, as the amount of PA produced in the early stage of anthrax invasion is relatively small. Graphene field-effect transistor (Gr−FET) biosensors are a new type of material for preparing biosensors, with the advantages of a short detection time and ultra-low detection limit. Methods: The effect of different concentrations of diluents on the affinity of PA monoclonal antibodies was determined via an ELISA experiment. Combined with the Debye equation, 0.01 × PBS solution was finally selected as the diluent for the experiment. Then, a PA monoclonal antibody was selected as the bio-recognition element to construct a Gr−FET device based on CVD-grown graphene, which was used to detect the concentration of PA while recording the response time, linear range, detection limit, and other parameters. Results: The experimental results showed that the biosensor could quickly detect PA, with a linear range of 10 fg/mL to 100 pg/mL and a detection limit of 10 fg/mL. In addition, the biosensor showed excellent specificity and repeatability. Conclusions: By constructing a Gr−FET device based on CVD-grown graphene and selecting a PA monoclonal antibody as the bio-recognition element, a highly sensitive, specific, and repeatable Gr−FET biosensor was successfully prepared for detecting extremely low concentrations of anthrax protective antigen (PA). This biosensor is expected to have a wide range of applications in clinical medicine and biological safety monitoring.

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“…Multiepitope antigens have been widely used for toxoplasmosis diagnosis; for example, synthetic multiepitope antigens are applied to detect anti-T. gondii IgG and IgM, and AMA1-SAG2-GRA1-ROP1 chimeric antigens are used to detect specific antibodies of human and mouse in early and chronic T. gondii infection [21]. Chimeric antigen technology has been developed for the serological diagnosis of Trypanosoma cruzi infection caused by another protozoan parasite, cutaneous anthrax caused by Bacillus anthracis, and others [22][23][24]. However, few studies have been conducted to evaluate chimeric antigens for serodiagnosis of T. gondii in pigs and to design an ELISA kit using synthetic antigens for the large-scale diagnosis of toxoplasmosis in pig farms and intensive industries.…”

Section: Introductionmentioning

confidence: 99%

Characterization and evaluation of a recombinant multiepitope peptide antigen MAG in the serological diagnosis of Toxoplasma gondii infection in pigs

et al. 2021

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Background Toxoplasmosis caused by Toxoplasma gondii is a serious disease threatening human and animal health. People can be infected with T. gondii by ingesting raw pork contaminated with cysts or oocysts. Serological test is a sensitive and specific method usually used for large-scale diagnosis of T. gondii infection in humans and animals (such as pigs). Commercial pig Toxoplasma antibody ELISA diagnostic kits are expensive, which limits their use; moreover, the wide antigen composition used in these diagnostic kits is still unclear and difficult to standardize. The multiepitope peptide antigen is a novel diagnostic marker, and it has potential to be developed into more accurate and inexpensive diagnostic kits. Methods The synthetic multiepitope antigen (MAG) cDNA encoding a protein with epitopes from five T. gondii-dominant antigens (SAG1, GRA1, ROP2, GRA4, and MIC3) was designed, synthesized, and expressed in Escherichia coli BL21 (DE3) strain. The recombinant protein was detected through western blot with pig anti-T. gondii-positive and -negative serum, and then IgG enzyme-linked immunosorbent assay (ELISA) named MAG-ELISA was designed. The MAG-ELISA was evaluated in terms of specificity, sensitivity, and stability. The MAG-ELISA was also compared with a commercial PrioCHECK®Toxoplasma Ab porcine ELISA (PrioCHECK ELISA). Finally, the trend of pig anti-T. gondii IgG levels after artificial infection with RH tachyzoites was evaluated using MAG-ELISA and two other ELISA methods (rMIC3-ELISA and PrioCHECK ELISA). Results MAG antigen could be specifically recognized by pig anti-T. gondii-positive but not -negative serum. MAG-ELISA showed high diagnostic performance in terms of specificity (88.6%) and sensitivity (79.1%). MAG-ELISA could be used for detecting anti-T. gondii IgG in the early stage of T. gondii infection in pigs (at least 7 days after artificial infection). Conclusions Our results suggest that MAG antigen can be applied to specifically recognize anti-T. gondii IgG in pig, and MAG-ELISA has the potential for large-scale screening tests of T. gondii infection in pig farms and intensive industries. Graphical abstract

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An ELISA using a recombinant chimera of protective antigen and lethal factor for serodiagnosis of cutaneous anthrax in India

Cited by 8 publications

References 33 publications

Dangerous Pathogens as a Potential Problem for Public Health

Dangerous Pathogens as a Potential Problem for Public Health

Research on Detection of Ultra-Low Concentration Anthrax Protective Antigen Using Graphene Field-Effect Transistor Biosensor

Characterization and evaluation of a recombinant multiepitope peptide antigen MAG in the serological diagnosis of Toxoplasma gondii infection in pigs

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