A technique for the purification of Mycobacterium paratuberculosis from the ileal mucosa of infected goats

The slow growth of M. paratuberculosis has hindered investigations of the antigenic composition of the orrganism and the development of species-specific antigen for serological detection of Johne's disase in goats. This paper describes a simple method for the isolation of large quantities of viable M. paratuberculosis from the intestinal mucosa of infected goats by a combination of trypsin digestion, deoxyribonuclease/lysozyme treatment and differential centrifugation. Purity was about 98.4% and yield between 10⁴–10⁸ bacteria/gm tissue.