Performance Estimation of Nested PCR-Based Assays for Direct Detection of Listeria monocytogenes in Artificially Contaminated Materials
Krzysztof Fiedoruk, Maria Lucyna Zaremba
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Department of Microbiology, Medical University of Białystok, A. Mickiewicza 2c, Białystok 15-222, Poland
Pol. J. Environ. Stud. 2010;19(2):293-299
Our study evaluated the sensitivity and repeatability of nested PCR-based assays for directly detecting L. monocytogenes in artificially contaminated human serum, pasteurized milk and physiological saline samples. The detection of the hlyA (267bp) and iap (371bp) gene fragments was compared. The logistic regression (logit model) was used to evaluate the probability of detection of L. monocytogenes at various contamination levels and to calculate the number of test repetitions required to reach necessary detection limits (e.g. 50%, 80%, 90%, and 95%). The reliable limit of detection for both genetic markers, ensuring ≥95% probability of detection, was established at 102 CFU/100μL.
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