| Real-time reverse transcriptase PCR for the rapid and sensitive detection of Salmonella typhimurium from pork. | |
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MedLine Citation:
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PMID: 20202337 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Reverse transcriptase PCR (RT-PCR) detects the presence of mRNA and has a greater potential for detecting viable pathogens than do DNA-based PCR assays, with improved speed and sensitivity compared with traditional methods. Our objective was to rapidly and sensitively detect Salmonella Typhimurium from pork within two 8-h work shifts using a SYBR Green I real-time RT-PCR (rt-RT-PCR) assay. Pork chop and sausage samples (25 g) were inoculated with 10(8) to 10(0) CFU of Salmonella Typhimurium and stomached in 225 ml of tetrathionate broth. Serial dilutions were spread plated on xylose lysine Tergitol 4 agar either immediately or after 10 h of selective preenrichment or preenrichment followed by 12 h of selective enrichment (for stressed cells) at 37 degrees C for standard cultural enumeration. RNA was extracted using the TRIzol method. The rt-RT-PCR assay was carried out in a Bio-Rad iCycler using a SYBR Green I one-step RT-PCR kit and Salmonella specific invA gene primers with an internal amplification control (IAC). The PCR was followed by melting temperature (T(m)) analysis to determine specific Salmonella invA (T(m) = 87.5 degrees C) and IAC (T(m) = 82 degrees C) products. Improved Salmonella detection up to 10(1) CFU/25 g of pork and 10(0) CFU/25 g of sausages was obtained after 10 h of enrichment within approximately 24 h. Even without enrichment, Salmonella could be detected from both pork chop and sausage at 10(6) CFU/25 g within 1 day. This robust rt-RT-PCR detects and confirms Salmonella in pork within approximately 24 h and thus is significantly faster than traditional methods that take >/=1 week. This assay shows promise for routine testing and monitoring of Salmonella by the pork industry. |
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Authors:
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Chayapa Techathuvanan; Frances Ann Draughon; Doris Helen D'Souza |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of food protection Volume: 73 ISSN: 0362-028X ISO Abbreviation: J. Food Prot. Publication Date: 2010 Mar |
Date Detail:
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Created Date: 2010-03-05 Completed Date: 2010-05-04 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 7703944 Medline TA: J Food Prot Country: United States |
Other Details:
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Languages: eng Pagination: 507-14 Citation Subset: IM |
Affiliation:
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Department of Food Science and Technology, University of Tennessee, Knoxville, Tennessee 37996-4591, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Bacterial Proteins Consumer Product Safety DNA, Bacterial / analysis Food Contamination / analysis* Food Microbiology Humans Meat / microbiology* Meat Products / microbiology RNA, Messenger / analysis Reverse Transcriptase Polymerase Chain Reaction* / standards Salmonella typhimurium / isolation & purification* Sensitivity and Specificity Swine Time Factors |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/DNA, Bacterial; 0/RNA, Messenger; 147652-43-5/invA protein, Bacteria |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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