Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2011 Aug 25:7:50.
doi: 10.1186/1746-6148-7-50.

Improved specificity for detection of Mycobacterium bovis in fresh tissues using IS6110 real-time PCR

Tyler C Thacker  1 , Beth Harris, Mitchell V Palmer, Wade R Waters
Affiliations

Improved specificity for detection of Mycobacterium bovis in fresh tissues using IS6110 real-time PCR

Tyler C Thacker et al. BMC Vet Res. .

Abstract

Background: Culture of M. bovis from diagnostic specimens is the gold standard for bovine tuberculosis diagnostics in the USA. Detection of M. bovis by PCR in tissue homogenates may provide a simple rapid method to complement bacterial culture. A significant impediment to PCR based assays on tissue homogenates is specificity since mycobacteria other than M. bovis may be associated with the tissues.

Results: Previously published IS6110 based PCR diagnostic assays, along with one developed in house, were tested against environmental mycobacteria commonly isolated from diagnostic tissues submitted to the National Veterinary Services Laboratory. A real-time PCR assay was developed (IS6110_T) that had increased specificity over other IS6110 based assays. Of the 13 non-tuberculous mycobacteria tested with IS6110_T only M. wolinskyi was positive. Thirty M. bovis infected tissue homogenates and 18 control tissues were used to evaluate the potential for the assay as a diagnostic test. In this small sample, IS6110_T detected 20/30 samples from M. bovis infected animals and 0/18 control tissues.

Conclusions: The IS6110_T assay provides a PCR based assay system that is compatible with current diagnostic protocols for the detection of M. bovis in the USA and compliments current testing strategies.

PubMed Disclaimer

References

    1. Plikaytis BB, Eisenach KD, Crawford JT, Shinnick TM. Differentiation of Mycobacterium tuberculosis and Mycobacterium bovis BCG by a polymerase chain reaction assay. Mol Cell Probes. 1991;5:215–219. doi: 10.1016/0890-8508(91)90043-J. - DOI - PubMed
    1. Eisenach KD, Cave MD, Bates JH, Crawford JT. Polymerase chain reaction amplification of a repetitive DNA sequence specific for Mycobacterium tuberculosis. J Infect Dis. 1990;161:977–981. doi: 10.1093/infdis/161.5.977. - DOI - PubMed
    1. Miller J, Jenny A, Rhyan J, Saari D, Suarez D. Detection of Mycobacterium bovis in formalin-fixed, paraffin-embedded tissues of cattle and elk by PCR amplification of an IS6110 sequence specific for Mycobacterium tuberculosis complex organisms. J Vet Diagn Invest. 1997;9:244–249. doi: 10.1177/104063879700900304. - DOI - PubMed
    1. Miller JM, Jenny AL, Payeur JB. Polymerase chain reaction detection of Mycobacterium tuberculosis complex and Mycobacterium avium organisms in formalin-fixed tissues from culture-negative ruminants. Vet Microbiol. 2002;87:15–23. doi: 10.1016/S0378-1135(02)00027-5. - DOI - PubMed
    1. Parra A, García N, García A, Lacombe A, Moreno F, Freire F, Moran J, Hermoso de Mendoza J. Development of a molecular diagnostic test applied to experimental abattoir surveillance on bovine tuberculosis. Vet Microbiol. 2008;127:315–324. doi: 10.1016/j.vetmic.2007.09.001. - DOI - PubMed

MeSH terms

LinkOut - more resources