Sero-reactivity prototype of secreted proteins of native ‘s 5’ vaccine strain to human sera positive for Mycobacterium paratuberculosis infection
Journal Title: Journal of Experimental Biology and Agricultural Sciences - Year 2017, Vol 5, Issue 0
Abstract
The zoonotic prospective of Mycobacterium avium subspecies paratuberculosis (MAP) has been debated for almost a century because of similarities between Johne’s disease (Paratuberculosis) in animals and Crohn’s disease (CD) in humans. Diagnosis of disease is hampered due to the extensive sharing of antigenic epitopes among MAP and other mycobacterial species. Therefore, this study is a preliminary work evaluated the reactivity pattern of secretory proteins harvested from native vaccine strain (MAP ‘S 5’) at different incubation time (8-12 weeks of growth). Analysis of secretory proteins was done by BioLogic LP chromatography system using Bio-ScaleTM Macroprep(R) High Q column (strong anion exchanger) and chromatogram showed six narrow protein peaks at 12 weeks by LP Data View v1.03 software (BioRad). Profile of secretory proteins harvested at different weeks was obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Immunoblotting showed strong reactivity of three secretory proteins commonly recognized (28, 34-38 and 42-45) with all three human sera positive for MAP infection. Additional 10 and 23-26 kDa proteins were recognized with two MAP infected human sera. Diagnostic potential of secretory proteins as a pool was evaluated using ‘Indirect ELISA test’. Results showed a lower sensitivity and 100% specificity with respect to semi-purified protoplasmic antigen (sPPA). Developing ‘ELISA based assay’ using these sero-reactive proteins will serve as backbone of future paratuberculosis control programs in the country. Earlier studies established that till today there are no universal diagnostic kits available for chronic diseases like CD. Therefore, study recommends use of secretory proteins as 'marker antigens' to design assay for the specific diagnosis of ‘active infection’ in human beings.
Authors and Affiliations
Saurabh Gupta, Shoor Vir Singh, A. K. Bhatia
Keywords
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- EP ID EP428051
- DOI 10.18006/2017.5(2).188.194
- Views 57
- Downloads 0
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