Effective DNA extraction method to improve detection of Mycobacterium avium subsp. paratuberculosis in bovine feces

Park, Hong-Tae; Shin, Min-Kyoung; Sung, Kyung Yong; Park, Hyun-Eui; Cho, Yong-Il; Yoo, Han Sang

doi:2014.54.1.55

Korean J Vet Res > Volume 54(1); 2014 > Article

Brief Communication

Korean Journal of Veterinary Research 2014;54(1):55-57.
DOI: https://doi.org/10.14405/kjvr.2014.54.1.55 Published online March 25, 2014.

Effective DNA extraction method to improve detection of Mycobacterium avium subsp. paratuberculosis in bovine feces

Hong-Tae Park¹, Min-Kyoung Shin¹, Kyung Yong Sung¹, Hyun-Eui Park¹, Yong-Il Cho², Han Sang Yoo¹

¹Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University
²Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration

Abstract

Paratuberculosis caused by Mycobacterium avium subsp. paratuberculosis (MAP) has extended latent periods of infection. Due to this property, difficulties in the detection of fecal shedder have been raised. A newly designed method for DNA extraction from fecal specimens, mGITC/SC was evaluated in terms of diagnostic efficiency. The detection limit of IS900 real-time PCR was about 50 MAP (1.5 cfu) in 250 mg of feces (6 cfu per g). Also, this DNA extraction method was faster and cheaper than that using commercial kit or other methods. Consequently, the mGITC/SC is an economical DNA extraction method that could be a useful tool for detecting MAP from fecal specimens.

Key Words: DNA extraction, feces, Mycobacterium avium subsp. paratuberculosis, real-time PCR