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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2-3
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pubmed:dateCreated |
1991-9-12
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pubmed:abstractText |
A sensitive non-isotopic assay for specific detection of reverse transcriptase (RT) of the human immunodeficiency virus type 1 (HIV-1) is described using 5-bromo-2'-deoxyuridine triphosphate (BrdUTP) instead of tritiated thymidine triphosphate. After the RT reaction the template primer is degraded by alkaline hydrolysis. Single-stranded poly.(BrdU) is detected in an immunoenzymometric assay using monoclonal anti-BrdU antibodies. The specificity of the assay is demonstrated by the isolation of RT from virus lysate by an insolubilised monoclonal anti-HIV-1 RT antibody prior to the RT reaction. Immunological RT binding leads to a tenfold increase in analytical sensitivity since substances inhibiting the RT reaction can be removed. This non-isotopic assay is some 30 times more sensitive than the classical radioisotopic RT assay. In terms of RT determination, however, there is a good correlation between these tests (r = 0.96). Several filtrations are no longer necessary to remove non-incorporated nucleotides. The test can be adapted to microtitre plates and hence is easy to automate.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyuracil Nucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/RNA-Directed DNA Polymerase,
http://linkedlifedata.com/resource/pubmed/chemical/Thymidine,
http://linkedlifedata.com/resource/pubmed/chemical/bromodeoxyuridine triphosphate
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pubmed:status |
MEDLINE
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pubmed:issn |
0166-0934
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
31
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
181-8
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:1713913-Antibodies, Monoclonal,
pubmed-meshheading:1713913-Deoxyuracil Nucleotides,
pubmed-meshheading:1713913-HIV-1,
pubmed-meshheading:1713913-Hydrolysis,
pubmed-meshheading:1713913-Immunoenzyme Techniques,
pubmed-meshheading:1713913-RNA-Directed DNA Polymerase,
pubmed-meshheading:1713913-Sensitivity and Specificity,
pubmed-meshheading:1713913-Templates, Genetic,
pubmed-meshheading:1713913-Thymidine
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pubmed:articleTitle |
A sensitive non-isotopic assay specific for HIV-1 associated reverse transcriptase.
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pubmed:affiliation |
Department of Medical Immunology, Medical School (Charité), Humboldt University Berlin, F.R.G.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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