Document Detail


Novel cost-efficient real-time PCR assays for detection and quantitation of Listeria monocytogenes.
MedLine Citation:
PMID:  21256878     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Listeriosis is a serious food-borne infection with mortality rates approaching 30%. Therefore, the rapid, cost-effective, and automated detection of Listeria monocytogenes throughout the food chain continues to be a major concern. Here we describe three novel quantitative real-time PCR assays for L. monocytogenes based on amplification of a target hlyA gene with SYBR Green I chemistry and hydrolysis probe (TaqMan MGB probe). In order to offer sensitive, rapid and robust tool of additional economical value the real-time PCR assays were designed and optimized to only 5μl-reactions. All assays were evaluated by using different non-reference Listeria strains isolated from various food matrices. Results demonstrated specificity to L. monocytogenes with accurate quantification over a dynamic range of 5-6 log units with R(2) higher than 0.98 and amplification efficiencies reaching above 92%. The detection and quantification limits were as low as 165 genome equivalents. Comparison of novel assays to commercially available TaqMan® Listeria monocytogenes Detection Kit and previously published studies revealed similar specificity, sensitivity and efficiency, but greater robustness and especially cost-efficiency in the view of smaller reaction volumes and continuous increase in sample throughput.
Authors:
Urban Traunšek; Nataša Toplak; Barbara Jeršek; Aleš Lapanje; Tamara Majstorovič; Minka Kovač
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-1-21
Journal Detail:
Title:  Journal of microbiological methods     Volume:  -     ISSN:  1872-8359     ISO Abbreviation:  -     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2011-1-31     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8306883     Medline TA:  J Microbiol Methods     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2011. Published by Elsevier B.V.
Affiliation:
Omega d.o.o., Dolinškova 8, Ljubljana, Slovenia.
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