PCR-based quantitation of low levels of HIV-1 DNA by using an external standard. NLM AIDSLINE Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.

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PCR-based quantitation of low levels of HIV-1 DNA by using an external standard.

Genet Anal Tech Appl. 1992 Aug;9(4):113-6. Unique Identifier : AIDSLINE MED/93112450
Arnold BL; Itakura K; Rossi JJ; Department of Molecular Genetics, City of Hope, Duarte, CA 91010.

Abstract: Beginning with 10(3)-10(5) molecules of a purified HIV-1 target sequence as a starting template, we have examined the effects of starting template concentration and cycle number on the amplification efficiency of the polymerase chain reaction. An external standard DNA sequence has been designed that when added to a DNA sample enables a determination of the starting concentration of HIV-1 target sequence in that sample of DNA. Varying ratios of external standard and target DNA sequences were amplified for 22 cycles. When the starting concentration of the external standard was within 50-fold of the starting concentration of the target, the amplifications of both sequences were proportional. These same results were obtained when the two templates were amplified in the presence of an excess of heterogeneous genomic DNA. Using this quantitative method, the number of starting target molecules in a DNA sample can be calculated to within a two-fold range of accuracy.
Keywords: Base Sequence DNA, Viral/*ANALYSIS HIV Long Terminal Repeat HIV-1/*CHEMISTRY Molecular Sequence Data Plasmids Polymerase Chain Reaction Reference Standards Support, Non-U.S. Gov't Templates JOURNAL ARTICLEKWDbasesequencedna,viral/KWDanalysishivlongterminalrepeathiv-1/KWDchemistrymolecularsequencedataplasmidspolymerasechainreactionreferencestandardssupport,non-uKWDsKWDgov'ttemplatesjournalarticle
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M9340784

Copyright © 1993 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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