Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol 16, Issue 1, 2014; Page No.(121-129)

EVALUATION OF ANTIBODIES RAISED AGAINST NATIVE AND RECOMBINANT ANTIGENS IN PLATE ELISA FORMAT FOR THE IMMUNOLOGICAL CHARACTERIZATION OF BURKHOLDERIA PSEUDOMALLEI CAUSATIVE ORGANISM OF MELIOIDOSIS

ARCHANA PRAKASH, SONIA ARORA, DURAIPANDIAN THAVASELVAM, ASHU KUMAR, ANITA BARUA AND K. SATHYASEELAN

Abstract

Melioidosis is an emerging infectious disease in India and the soil dwelling Gram-negative pathogen Burkholderia pseudomallei is the causative organism for Melioidosis. The disease is endemic in Southeast Asia and northern Australia with highest percentage of melioidosis cases reported by Thailand. This disease is increasingly becoming an emerging disease in mostly coastal regions of India. In the present study antibodies raised against several native and recombinant antigens of Burkholderia pseudomallei was evaluated in plate ELISA format for the immunological characterization of clinical and environmental isolates. In all sixty nine whole cell native antigens of Burkholderia pseudomallei strains, Burkholderia related and other species were tested with rabbit polyclonal antibodies raised against sonicated antigen of B. pseudomallei NCTC10274, mice anti-recombinant GroEL protein antigen, mice anti-cell envelope protein antigen, monoclonal antibodies raised against whole cell sonicated antigen and human Melioidosis sera from culture positive clinical case as positive control. The polyclonal antibodies raised against sonicated antigen of B. pseudomallei reference strain was found cross reactive with 20 other bacterial species and polyclonal antibodies raised against cell envelope antigen reacted with 5 bacterial species. The polyclonal antibodies raised against recombinant GroEL protein cross reacted with 8 bacterial species and the pooled human culture positive sera reacted with six bacterial species. All the polyclonal antibodies tested although had positive reaction with all the reference strains, clinical and environmental isolates of B. pseudomallei but cross reacted with other bacteria. The two monoclonal antibodies tested reacted selectively with the B. pseudomallei isolates and no reaction was observed with any of the other bacterial species tested. These results will be very useful in not only development of immuno assay for detection but also for rapid characterization of isolates.

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