Development & validation of a quantitative anti-protective antigen IgG enzyme linked immunosorbent assay for serodiagnosis of cutaneous anthrax.

Abstract

BACKGROUND & OBJECTIVES:

Anthrax caused by Bacillus anthracis is primarily a disease of herbivorous animals, although several mammals are vulnerable to it. ELISA is the most widely accepted serodiagnostic assay for large scale surveillance of cutaneous anthrax. The aims of this study were to develop and evaluate a quantitative ELISA for determination of IgG antibodies against B. anthracis protective antigen (PA) in human cutaneous anthrax cases.

METHODS:

Quantitative ELISA was developed using the recombinant PA for coating and standard reference serum AVR801 for quantification. A total of 116 human test and control serum samples were used in the study. The assay was evaluated for its precision, accuracy and linearity.

RESULTS:

The minimum detection limit and lower limit of quantification of the assay for anti-PA IgG were 3.2 and 4 µg/ml, respectively. The serum samples collected from the anthrax infected patients were found to have anti-PA IgG concentrations of 5.2 to 166.3 µg/ml. The intra-assay precision per cent CV within an assay and within an operator ranged from 0.99 to 7.4 per cent and 1.7 to 3.9 per cent, respectively. The accuracy of the assay was high with a per cent error of 6.5 - 24.1 per cent. The described assay was found to be linear between the range of 4 to 80 ng/ml (R [2] = 0.9982; slope = 0.9186; intercept = 0.1108).

INTERPRETATION & CONCLUSIONS:

The results suggested that the developed assay could be a useful tool for quantification of anti-PA IgG response in human after anthrax infection or vaccination.

• Authors: N Ghosh, D Gunti, H Lukka, B R Reddy, Jyothi Padmaja, A K Goel
• Journal: The Indian journal of medical research (Indian J Med Res) Vol. 142 Issue 2 Pg. 196-204 (Aug 2015) ISSN: 0971-5916 [Print] India
• PMID: 26354217 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Anti-Idiotypic
• Antibodies, Bacterial
• Antigens, Bacterial
• Immunoglobulin G
• anti-IgG

Topics

• Anthrax (blood, immunology)
• Antibodies, Anti-Idiotypic (immunology, isolation & purification)
• Antibodies, Bacterial (blood, immunology)
• Antigens, Bacterial (immunology)
• Bacillus anthracis (immunology, isolation & purification, pathogenicity)
• Enzyme-Linked Immunosorbent Assay (methods)
• Humans
• Immunoglobulin G (blood, immunology, isolation & purification)
• Serologic Tests
• Skin Diseases, Bacterial (blood, immunology)