Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
9
pubmed:dateCreated
2007-5-7
pubmed:abstractText
We developed a 12-h Salmonella detection method, based on 8 h of preenrichment, followed by automated DNA extraction and a sensitive real-time PCR. The method was optimized to obtain the highest possible yield of cells and DNA. The growth of different Salmonella strains in various preenrichment media and the effects of adding growth-promoting and selective reagents were explored, taking into account their PCR compatibility. The effects of (i) analyzing larger volumes (1 to 5 ml) from preenriched samples and introducing wash steps prior to DNA extraction, (ii) regulating the amount of paramagnetic particles (increasing it from 60 to 90 microl) in the DNA extraction, (iii) eluting the DNA in reduced volumes (25 or 50 microl rather than 100 microl), and (iv) increasing the PCR template volume (from 5 to 20 microl) were investigated. After 8 h of preenrichment, buffered peptone water yielded the highest number of salmonellae. When analyzing minced meat samples, positive effects of increasing the initial sampling volume from 1 to 5 ml and increasing the amount of paramagnetic particles to 90 microl were observed. However, washing the pellet and eluting the DNA in reduced volumes (25 and 50 microl) had no positive effects and resulted in decreased reproducibility. Increasing the amount of PCR template DNA from 5 to 20 mul improved the threshold cycle value by approximately 2. The improved 12-h PCR method was successfully compared to a reference culture method with 100 minced meat and poultry samples, with a relative accuracy of 99%, a relative sensitivity of 98%, and a relative specificity of 100%.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-10645983, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-11056764, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-11108192, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-11773092, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-11849490, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-12270264, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-12512356, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-12581847, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-14500016, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-14633109, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-15036370, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-15574899, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-16145116, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-16364478, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-16564585, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-16690151, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-16885299, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-16943057, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-1908453, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-334073, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-3932291, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-7929768, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-8722189, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-9023957, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-9105933, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-9717315, http://linkedlifedata.com/resource/pubmed/commentcorrection/17351094-9721644
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0099-2240
pubmed:author
pubmed:issnType
Print
pubmed:volume
73
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3040-8
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2007
pubmed:articleTitle
Optimization of a 12-hour TaqMan PCR-based method for detection of Salmonella bacteria in meat.
pubmed:affiliation
National Food Institute, Bülowsvej 27, DK-1790 Copenhagen, Denmark.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't