Linked Life Data

8789009

Source:http://linkedlifedata.com/resource/pubmed/id/8789009

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1996-10-21
pubmed:abstractText
Quantitative tests for human immunodeficiency virus type 1 (HIV-1) RNA in plasma and proviral DNA in peripheral blood mononuclear cells (PBMC) provide valuable information on the status of HIV-1 infection. This paper describes tests that were carried out with commercially available materials and an enzyme-linked immunosorbent assay reader for detecting spectrophotometric changes. Samples consisted of 100 microliters of plasma or 200,000 PBMC. The procedure involved sample preparation, PCR-based amplification with the primer pair SK39 (biotinylated at the 5' end) and SK38, hybridization of the cDNA PCR product to an RNA probe, capture of the RNA-DNA hybrid on a solid phase by means of strepavidin, binding to an alkaline phosphatase-conjugated antibody directed against RNA-DNA hybrids, and incubation with p-nitrophenylphosphate. Spectrophotometric changes were recorded at four intervals over a period of 20 h. The inclusion of HIV-1 RNA or proviral DNA standards in each run was an integral part of the procedure. The dynamic ranges afforded by these assays--500 to 1 million RNA copies per ml and 10 to 5,000 proviral DNA copies per 10(6) PBMC--were applicable to most plasma specimens and to all PBMC specimens from HIV-1-infected patients. Correlations of log-transformed HIV-1 RNA and proviral DNA concentrations with those found by reference methods were, respectively, 0.88 and 0.80. The between-run coefficients of variation for the detection method were < or = 25% (range, 9.1 to 24.7) and < or = 15% (range, 10.9 to 15.1), respectively, for HIV-1 RNA and proviral DNA. The reproducibility of the overall procedure for HIV-1 RNA in plasma (including sample preparation, amplification, and detection) was given by a duplicate standard deviation of log10 copies per ml of 0.11. Thus, the method was sufficiently precise to allow the detection of fourfold changes in plasma HIV-1 RNA concentrations, with a power of 0.95.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-1572969, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-2440339, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-3014036, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-7693821, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-7699034, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-7718186, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-7816094, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-7903317, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-7915748, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-8096089, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-8150937, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-8150964, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-8253949, http://linkedlifedata.com/resource/pubmed/commentcorrection/8789009-8308102
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0095-1137
pubmed:author
pubmed:issnType
Print
pubmed:volume
34
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
329-33
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1996
pubmed:articleTitle
Application of a commercial kit for detection of PCR products to quantification of human immunodeficiency virus type 1 RNA and proviral DNA.
pubmed:affiliation
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030, USA. lin@bcm.tmc.edu
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't