Journal
FOOD CONTROL
Volume 25, Issue 1, Pages 117-124Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.foodcont.2011.10.025
Keywords
Listeria monocytogenes; Real-time reverse-transcriptase PCR; Detection; RNA; Chilled pork
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Funding
- Ministry of Science and Technology of China [2009DFA 31770]
- National Natural Science Foundation of China [31071614]
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The objective of this study was to develop an RNA-dependent real-time reverse-transcriptase PCR (real-time RT-PCR) method for the detection of Listeria monocytogenes in chilled pork without the need for pre-enrichment steps, and the soundness of the method was simultaneously validated and evaluated by DNA-based real-time PCR and traditional culture methods. For specificity testing, a lack of amplification signals and no Tm peak at similar to 78.37 degrees C were obtained from any of 41 other bacterial strains associated with meat species under the conditions used. The R-2 and efficiency of standard curves constructed by ten-fold serial dilutions of pure L monocytogenes were respectively 0.995 and 90.1%; lower than that of the DNA-based assay. The detection limit was up to 10(0) cfu/mL in both pure culture and in artificially contaminated chilled pork samples. Quantitative detection showed that the RNA-based assay obtained relatively accurate results when samples had undergone treatments (such as high pressure), but without treatment, the results showed a slight deviation compared with plate counts. The RNA-dependent real-time RT-PCR method developed in this study was found to be rapid and sensitive and should be useful for reliable detection of viable L. rnonocytogenes in chilled pork, especially for pre-treated samples. However, this method cannot be recommended to accurately quantify L monocytogenes, but only to show the presence of live cells and approximately predict its level of contamination. (c) 2011 Elsevier Ltd. All rights reserved.
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