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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
7
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pubmed:dateCreated |
1992-8-20
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pubmed:abstractText |
In order to identify methicillin-resistant staphylococci from clinical sources with ease and reliability, enzymatic detection of polymerase chain reaction (ED-PCR) was applied. ED-PCR is based on the capture of amplified products via biotin-streptavidin affinity and the detection of an incorporated hapten in amplified products with an enzyme-linked antibody. In order to identify methicillin-resistant staphylococci of all species, a 150-bp fragment of the mecA gene was targeted for ED-PCR. After PCR was performed with a pair of biotin and dinitrophenol 5'-labeled primers, the reaction mixture was applied to a microtiter well precoated with streptavidin. Thereafter, bound PCR products were detected colorimetrically with alkaline phosphatase-conjugated anti-dinitrophenol antibody. The extraction of DNA from staphylococcal cells for PCR was simplified so that it could be performed within one tube. The total assay, including PCR, took less than 3 h. The sensitivity of mecA gene detection ranged from greater than 5 x 10(2) CFU per tube for Staphylococcus aureus to greater than 5 x 10(3) CFU per tube for Staphylococcus epidermidis. Genotyping results obtained by ED-PCR of 161 tested strains from the colonies (97 strains of S. aureus and 64 strains of coagulase-negative staphylococci) were compared with the phenotypic susceptibilities of the strains to oxacillin. The results of ED-PCR showed excellent agreement with the MICs of oxacillin with very few exceptions; only one strain of S. aureus and two strains of coagulase-negative staphylococci were found to possess the mecA gene, which was discrepant with their phenotypes. Fifty-five blood culture samples were also tested by ED-PCR. For staphylococcal isolates in 33 of the cultures, oxacillin MICs were >4 microgram/ml; 31 of the 33 staphylococcal isolates were determined by ED-PCR to be mecA gene positive. These results suggest that ED-PCR can be used with reasonable confidence in the clinical microbiological laboratory.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-1691614,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-1939577,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2039210,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2048117,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2069369,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2227446,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2285281,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2285284,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2564067,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2610497,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2675760,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2708325,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2729937,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2817861,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-2903715,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3221195,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3266728,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3305073,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3488015,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3499861,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3638304,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3848293,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3848294,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-3878127,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1629327-6563036
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0095-1137
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
30
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pubmed:geneSymbol |
mecA
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1728-33
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:1629327-Alkaline Phosphatase,
pubmed-meshheading:1629327-Base Sequence,
pubmed-meshheading:1629327-Drug Resistance, Microbial,
pubmed-meshheading:1629327-Genes, Bacterial,
pubmed-meshheading:1629327-Methicillin Resistance,
pubmed-meshheading:1629327-Molecular Sequence Data,
pubmed-meshheading:1629327-Oxacillin,
pubmed-meshheading:1629327-Polymerase Chain Reaction,
pubmed-meshheading:1629327-Sensitivity and Specificity,
pubmed-meshheading:1629327-Staphylococcus
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pubmed:year |
1992
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pubmed:articleTitle |
Rapid detection of the mecA gene in methicillin-resistant staphylococci by enzymatic detection of polymerase chain reaction products.
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pubmed:affiliation |
Department of Clinical Pathology, School of Medicine, Teikyo University, Tokyo, Japan.
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pubmed:publicationType |
Journal Article
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