Sensitive detection of Mycobacterium avium subsp. paratuberculosis in bovine semen by real-time PCR

Journal of Applied Microbiology
D HerthnekG Bölske

Abstract

To develop a fast and sensitive protocol for detection of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine semen and to make a critical evaluation of the analytical sensitivity. Processed semen was spiked with known amounts of MAP. Semen from different bulls as well as semen of different dilutions was tested. The samples were treated with lysing agents and beadbeating and the DNA was extracted with phenol and chloroform. Real-time PCR with a fluorescent probe targeting the insertion element IS900 detected as few as 10 organisms per sample of 100 mul semen. PCR-inhibition was monitored by inclusion of an internal control. Pre-treatment with immunomagnetic separation was also evaluated, but was not shown to improve the overall sensitivity. Real-time PCR is a sensitive method for detection of MAP in bovine semen. Lysis by mechanical disruption followed by phenol and chloroform extraction efficiently isolated DNA and removed PCR-inhibitors. The high sensitivity of the applied method allows reliable testing of bovine semen used for artificial insemination to prevent the spread of Johne's disease, caused by MAP.

References

Oct 1, 1991·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·D L WhippleJ L Jarnagin
Apr 1, 1995·Letters in Applied Microbiology·J H DickinsonK A Grant
Sep 3, 1998·Applied and Environmental Microbiology·I R GrantM T Rowe
Nov 18, 1998·Journal of Clinical Microbiology·G Zhang, A Weintraub
Jul 11, 2000·Canadian Journal of Gastroenterology = Journal Canadien De Gastroenterologie·J Hermon-TaylorN Sumar
Apr 6, 2001·Revue Scientifique Et Technique·D J Kennedy, G Benedictus
Jan 5, 2002·Journal of Food Protection·J E Harris, A M Lammerding
Jan 16, 2002·Australian Veterinary Journal·J Eppleston, R J Whittington
May 15, 2002·FEMS Microbiology Letters·Stina EnglundKarl Erik Johansson
Dec 19, 2002·Veterinary Microbiology·L ManterolaI López-Goñi
May 18, 2004·Veterinary Microbiology·Elke VansnickFrançoise Portaels

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Citations

Oct 12, 2012·Tropical Animal Health and Production·Mohamed SalemGeorge Baljer
Oct 18, 2011·Applied and Environmental Microbiology·Natalia ElguezabalRamón A Juste
Nov 26, 2011·Journal of Crohn's & Colitis·Herbert J Van Kruiningen
Mar 4, 2011·Research in Veterinary Science·Elisa MaioChristian Gortázar
Jul 28, 2006·Reproduction in Domestic Animals = Zuchthygiene·L de RuighJ S Merton
May 3, 2008·International Journal of Food Microbiology·W J MeadusL Saucier
Dec 5, 2019·Veterinary Sciences·Karianne Lievaart-PetersonAd P Koets
Jan 1, 2011·European Journal of Wildlife Research·Raquel SobrinoChristian Gortazar

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