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Correlation between Herrold egg yolk medium culture and real-time quantitative polymerase chain reaction results for Mycobacterium avium subspecies paratuberculosis in pooled fecal and environmental samples

Permanent URL:
http://handle.nal.usda.gov/10113/48587
Abstract:
Real-time quantitative polymerase chain reaction (qPCR) testing for Mycobacterium avium subspecies paratuberculosis (MAP) in fecal samples is a rapid alternative to culture on Herrold egg yolk medium (HEYM), the traditional antemortem reference test for MAP. Although the sensitivity and specificity of these 2 tests have been estimated based on dichotomized test results, the correlation between real-time qPCR threshold cycle (Ct) values and colony-forming units (CFU) on HEYM for fresh and thawed samples has not been evaluated. The objectives of the present study were to estimate the correlation and association between Ct and CFU in fresh and thawed pooled fecal and environmental samples. Results of HEYM culture of 1,997 pooled fecal samples from cows in 14 herds, and 802 environmental samples from 109 dairies nationwide were negatively (inversely) correlated with their respective real-time qPCR results. The Spearman's rank correlation between Ct and CFU was good (–0.66) in fresh and thawed pooled fecal samples, and excellent (–0.76) and good (–0.61) in fresh and thawed environmental samples, respectively. The correlation varied from good (–0.53) to excellent (–0.90) depending on the number of samples in a fecal pool. Truncated regression models indicated a significant negative association between Ct and CFU in fecal pools and environmental samples. The use of real-time qPCR instead of HEYM can yield rapid, quantitative estimates of MAP load and allow for incorporation of real-time qPCR results of pooled and environmental samples in testing strategies to identify dairy cow groups with the highest MAP shedding.
Author(s):
Aly, Sharif S. , Mangold, Beverly L. , Whitlock, Robert H. , Sweeney, Raymond W. , Anderson, Randall J. , Jiang, Jiming , Schukken, Ynte H. , Hovingh, Ernest , Wolfgang, David , Van Kessel, Jo Ann S. , Karns, Jeffrey S. , Lombard, Jason E. , Smith, Julia M. , Gardner, Ian A.
Subject(s):
Mycobacterium avium subsp. paratuberculosis , animal pathogenic bacteria , feces , sampling , polymerase chain reaction , pathogen identification , detection , culture media , egg yolk , cultured cells , reference standards , accuracy , precision , disease surveillance , rapid methods , quantitative analysis
Format:
p. 677-683.
Note:
Includes references
Source:
Journal of veterinary diagnostic investigation 2010 Sept., v. 22, no. 5
Language:
English
Year:
2010
Collection:
Journal Articles, USDA Authors, Peer-Reviewed
File:
Download [PDF File]
Rights:
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.