Advances in Animal and Veterinary Sciences

Research Article
Adv. Anim. Vet. Sci. 3 (6): 347 - 353

Immunoreactivity to Culture Filtrate Proteins of Mycobacterium avium Subspecies paratuberculosis in Naturally Infected Goat and Sheep Sera

Saurabh Gupta1, 2, Kundan Kumar Chaubey1, Shoor Vir Singh1*, Ashok Kumar Bhatia2, Naveen Kumar1, Anjana Goel2, Tarun Kumar Sachan1, Krishan Dutta Rawat1, Jagdip Singh Sohal3, Kuldeep Dhama4

1Microbiology Laboratory, Animal Health Division, Central Institute for Research on Goats, Makhdoom, PO-Farah, Mathura, Uttar Pradesh, India; 2Department of Biotechnology, GLA University, Chaumuhan, Mathura, Uttar Pradesh, India; 3Amity Institute of Microbial Technology, Amity University Rajasthan, Jaipur, India; 4Division of Pathology, Indian Veterinary Research Institute, Izatnagar, Bareilly, Uttar Pradesh, India.

*Correspondence | Shoor Vir Singh, Central Institute for Research on Goats, Makhdoom, Mathura, Uttar Pradesh, India; Email:

Mycobacterium avium subspecies paratuberculosis (MAP), the cause of granulomatous chronic enteritis in ruminants (Johne’s disease) is under reported due to difficulties in diagnosing pre-clinical cases. Compromised specificity is a problem due to extensive sharing of antigens /epitopes among MAP and other mycobacterial strains. Culture filtrate (CF) proteins profile of native ‘Indian Bison Type’ strain of MAP was studied at different harvesting times (2-8 weeks of growth) in Middlebrook 7H9 medium supplemented with ADC, PANTA antibiotics and mycobactin J. Analysis of harvested CF proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the greater part of CF proteins had molecular masses (<70 kDa) as 14, 19, 26, 34-41, 52-55, and 70 kDa. Immunoblotting showed reactivity of CF proteins commonly recognised (28, 34-36, 38-42, 45, and 56 kDa) with all four MAP infected goat and sheep sera at 2-8 weeks of growth. Collectively, these immunoreactive MAP CF proteins could be the potential targets for developing diagnostics against Johne’s disease with improved sensitivity and high specificity instead of whole cell sonicated crude protoplasmic extracts (PPA).

Keywords | Johne’s disease, Mycobacterium avium subspecies paratuberculosis, Culture filtrate, SDS-PAGE, Immunoblotting

Pakistan Journal of Zoology


Pakistan J. Zool., Vol. 55, Iss. 6, pp. 2501-3000


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