Korean J Vet Res.  2014 Mar;54(1):55-57.

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

Affiliations
  • 1Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. yoohs@snu.ac.kr
  • 2Department of Animal Resources Development, National Institute of Animal Science, Rural Development Administration, Cheonan 331-801, Korea.
  • 3Institute of Green Bio Science and Technology, Seoul National University, Pyeongchang 232-916, Korea.

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.

Keyword

DNA extraction; feces; Mycobacterium avium subsp. paratuberculosis; real-time PCR

MeSH Terms

Animals
DNA*
Feces*
Limit of Detection
Mycobacterium avium subsp. paratuberculosis*
Mycobacterium avium*
Mycobacterium*
Paratuberculosis
Real-Time Polymerase Chain Reaction
DNA
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