1. Application of antibodies to recombinant heat shock protein 70 in immunohistochemical diagnosis of mycobacterium avium subspecies paratuberculosis in tissues of naturally infected cattle
- Author
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Moses Okee, David P. Kateete, Julius Boniface Okuni, Anna Nanteza, Moses Joloba, and Lonzy Ojok
- Subjects
0301 basic medicine ,040301 veterinary sciences ,law.invention ,0403 veterinary science ,03 medical and health sciences ,law ,Heat shock protein ,HSP70 ,General Veterinary ,biology ,Johne’s disease ,Research ,04 agricultural and veterinary sciences ,biology.organism_classification ,Mycobacterium avium subspecies paratuberculosis ,Virology ,Primary and secondary antibodies ,Immunohistochemistry ,Hsp70 ,030104 developmental biology ,Polyclonal antibodies ,Recombinant DNA ,biology.protein ,Antibody - Abstract
Background Detection of Mycobacterium avium subspecies paratuberculosis (MAP) infection is key to the control of Johne’s disease. Immunohistochemistry is one of the methods of detection of MAP infection in tissues. However, unavailability of commercial antibodies that can detect the organism is a limiting factor for the use of immunohistochemistry. This study was aimed at developing an immunohistochemistry method to diagnose MAP in infected tissues using antibodies against MAP recombinant heat shock protein 70kd. Results MAP Heat shock protein 70 gene was amplified and cloned into an expression vector, Champion pET-SUMO, then expressed in E coli, purified and used to produce polyclonal rabbit antibodies against the Heat shock protein. Immunohistochemistry was performed in 35 MAP infected tissues with anti-HSP70 polyclonal antibodies. All 35 MAP infected tissues were positive for MAP within macrophages, epithelioid cells and giant cells either in clumps or singly as individual bacilli. No positive staining was seen in the three uninfected normal tissues and in MAP infected tissues where primary antibodies were substituted with PBS or pre-immune serum from the same rabbit. Conclusion Anti-HSP70 produced in this study offers an opportunity for improved diagnosis, screening of MAP in animal tissues and in studies on the pathogenesis of MAP Electronic supplementary material The online version of this article (doi:10.1186/s13620-017-0088-7) contains supplementary material, which is available to authorized users.
- Published
- 2017