A Rapid Flow through Membrane Enzyme Linked Immunosorbent Assay for Bacillus anthracis using Surface Array Protein as a Biomarker

Nidhi Puranik; Manoj Kumar; N. Tripathi; V. Pal; A. K. Goel

doi:10.14429/dsj.69.13730

Nidhi Puranik DRDO-Defence Research and Development Establishment, Gwalior - 474 002
Manoj Kumar DRDO-Defence Research and Development Establishment, Gwalior - 474 002
N. Tripathi DRDO-Defence Research and Development Establishment, Gwalior - 474 002
V. Pal DRDO-Defence Research and Development Establishment, Gwalior - 474 002
A. K. Goel DRDO-Defence Research and Development Establishment, Gwalior - 474 002

DOI: https://doi.org/10.14429/dsj.69.13730

Keywords: Bacillus anthracis, ELISA, Polyclonal antibody, Surface array protein

Abstract

Anthrax, caused by Bacillus anthracis is an important disease of biowarfare and public health importance. It is imperative to develop a simple system which can detect and differentiate B. anthracis from other closely related species. The surface array protein (Sap), which is secreted during the early growth phase of bacteria can be an important biomarker for detection of B. anthracis. In the present study, we have developed a rapid flow through membrane ELISA for detection of B. anthracis. Polyclonal antibodies were used to develop a sandwich plate ELISA, which could detect 3.9 ng/ml of recombinant Sap. B. anthracis bacteria grown in culture broth could be detected after 5 h of growth. Finally, a rapid flow through membrane ELISA was developed which can be accomplished just within 2 minutes, instead of 3-4 h as required in sandwich plate ELISA. The results established that the developed flow through membrane ELISA may be used for detection of B. anthracis. The proposed method is rapid, safe and user friendly for detection of B. anthracis culture.

Author Biographies

Nidhi Puranik, DRDO-Defence Research and Development Establishment, Gwalior - 474 002

Ms Nidhi Puranik received the Master from Devi Ahilya Vishwavidyalaya (DAVV), Indore, M.P., India in 2009. Currently working as SRF at Defence Research & Development Establishment and pursuing PhD from Bharathiar University, Coimbatore. She is working on development of rapid immunological systems for detection and diagnosis of anthrax from environmental and clinical samples. She performed the experiments and wrote the
manuscript in this study.

Manoj Kumar, DRDO-Defence Research and Development Establishment, Gwalior - 474 002

Mr Manoj Kumar received Master from Jiwaji University, Gwalior, India in 2011. Currently working as SRF at Defence Research & Development Establishment and pursuing PhD from Bharathiar University, Coimbatore. He is working on identification and characterisation of immunodominant antigen/s: evaluation as subunit vaccine candidate/s in a mouse model against Bacillus anthracis.

N. Tripathi, DRDO-Defence Research and Development Establishment, Gwalior - 474 002

Dr Nagesh Tripathi received his PhD (Chemical Engineering) from National Institute of Technology, Rourkela. Presently, he is scientist ‘D’ at the Defence Research and Development Establishment, Gwalior. His research interest includes scale up of biomolecules including recombinant proteins, activated carbon spheres and development of chemical protective suit.

V. Pal, DRDO-Defence Research and Development Establishment, Gwalior - 474 002

Dr Vijai Pal did his Master’s in Biotechnology from CCS Haryana Agricultural University Hisar in 2000 and PhD from Jiwaji University, Gwalior in 2016. He joined as Scientist ‘B’ at Defence Food Research Laboratory, Mysore in 2001. Presently, he is working as Scientist ‘E’ at Defence Research and Development Establishment, Gwalior on development of diagnostic/detection systems for biothreat agents. He has published more than 23 research papers in Indian and International Journals, besides
one book and has filed one Indian Patent.

A. K. Goel, DRDO-Defence Research and Development Establishment, Gwalior - 474 002

Dr Ajay Kumar Goel received his PhD (Microbiology) from CCS Haryana Agricultural University, Hisar, in 1999. Currently working as a Scientist ‘F’ and Head, Bioprocess Technology Division, Defence Research and Development Establishment, Gwalior. He has more than 100 research papers, 6 patents, radio talks, books and several overseas presentations to his credit. His current research interest includes development of detection and protection systems for potential biothreat agents.