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United States Department of Agriculture

Agricultural Research Service

Title: Evaluation of the Reproducibility and Accuracy of a Practical Pcr Assay for Rapid Detection and Differentiation of Strains of the Mycobacterium Avium Complex

item Ellingson, Jay
item Stabel, Judith
item Frothingham, R - VA MEDICAL CENTER & DUKE
item Miller, Janice

Submitted to: American Society for Microbiology
Publication Type: Abstract Only
Publication Acceptance Date: May 18, 1998
Publication Date: N/A

Technical Abstract: The Mycobacterium avium complex (MAC) includes the closely related species M. avium, M. intracellulare, and M. paratuberculosis. Members of MAC are known pathogens of animals and humans. Rapid identification and differentiation of MAC species is important for diagnosis and treatment of infections by these pathogens. Oligonucleotide primers derived from the 16S rRNA (MAC) sequence, insertion elements IS900 (M. paratuberculosis), IS901 (M. avium), IS1245 (MAC), and the hspX gene, a gene unique to M. paratuberculosis, were synthesized and used in a polymerase chain reaction (PCR) assay. For simplicity, different colored tubes were used for each primer set reaction mix. To determine the repeatability and accuracy of the PCR assay as a potential diagnostic tool, MAC isolates (n = 100) from a pigeon, cattle, humans, and a primate were screened by the PCR panel assay by one individual. For evaluation of repeatability, samples were numbered, ,given to a different individual as unknowns and identified and differentiated by PCR using the panel of color coded primer set reaction tubes. In this study, the internal repeatability of the PCR results was 100% (100/100). The experiments indicate that this rapid PCR method using the panel of color coded primer set reaction tubes can identify and differentiate closely related members of MAC with ~ 95% accuracy. The simplicity of this PCR method could be beneficial to laboratories that test for members of MAC.

Last Modified: 4/21/2015