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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
12
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pubmed:dateCreated |
1996-3-27
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pubmed:abstractText |
Two quantitative PCR methods with our nonisotopic enzyme-linked oligosorbent assay (ELOSA) in microtiter plate format were developed for quantitation of human immunodeficiency virus type 1 (HIV-1). Quantitative competitive PCR (QC-PCR) was based on the coamplification of the wild-type nef region with a mimic competitive nef gene template carrying mutations in the capture region. Correlation of wild-type HIV-1 nef DNA to mimic template copy number permitted quantitation of HIV-1 copy numbers in the range of 20 to 2,000 copies per micrograms of DNA. Internally controlled PCR (IC-PCR) was based on coamplification of the nef region and the ras gene as an internal endogenous standard. Correlation to known amounts of HIV-1 DNA permitted quantitation by IC-PCR of HIV-1 copy numbers in the range of 10 to 2,000 copies per microgram of DNA. QC- and IC-PCR-ELOSA were performed on a panel of 53 seropositive patients and 12 seronegative controls. The methods showed similar coefficients of variation below 24%. Quantitations by QC- and IC-PCR-ELOSA were identical for 77% of patient samples. The copy level ranged between 443 +/- 156 and 21,453 +/- 13,511 copies per 10(5) CD4 cells for asymptomatic and AIDS patients, respectively. The simplicity and reliability of QC- and IC-PCR-ELOSA methods make them appropriate for routine laboratory use in the quantitation of viral and bacterial DNAs.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-1333488,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-1347227,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-1353103,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-1357296,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-1472386,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-1490169,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-1909145,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-2127682,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-2229398,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-2230230,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-2281864,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-2296085,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-2463307,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-3387214,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-7541215,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-7679276,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-7680263,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-7786582,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-7909225,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-8150972,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-8161441,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-8161443,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-8314984,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-8346024,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-8424881,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8586703-8441670
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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 |
Dec
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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 |
33
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
3201-8
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:8586703-Acquired Immunodeficiency Syndrome,
pubmed-meshheading:8586703-Base Sequence,
pubmed-meshheading:8586703-DNA, Viral,
pubmed-meshheading:8586703-DNA Primers,
pubmed-meshheading:8586703-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:8586703-Evaluation Studies as Topic,
pubmed-meshheading:8586703-Genes, nef,
pubmed-meshheading:8586703-HIV Infections,
pubmed-meshheading:8586703-HIV-1,
pubmed-meshheading:8586703-Humans,
pubmed-meshheading:8586703-Indicators and Reagents,
pubmed-meshheading:8586703-Molecular Sequence Data,
pubmed-meshheading:8586703-Polymerase Chain Reaction,
pubmed-meshheading:8586703-Quality Control,
pubmed-meshheading:8586703-Reproducibility of Results
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pubmed:year |
1995
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pubmed:articleTitle |
Quantitation of human immunodeficiency virus type 1 DNA by two PCR procedures coupled with enzyme-linked oligosorbent assay.
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pubmed:affiliation |
Unité Mixte de Recherche 103 Centre National de la Recherche Scientifique-bioMérieux, Ecole Normale Supérieure de Lyon, France.
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pubmed:publicationType |
Journal Article,
Comparative Study
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