1991
DOI: 10.1182/blood.v77.4.879.879
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Rapid simultaneous detection of multiple retroviral DNA sequences using the polymerase chain reaction and capillary DNA chromatography

Abstract: The polymerase chain reaction (PCR) technique is a powerful new tool for amplifying target DNA, thus allowing for sensitive detection of specific nucleic acid sequences. One important potential use of PCR involves screening the donated blood supply for transfusion-transmitted viruses. Realization of this goal has been limited by (1) the requirement for multiple, discrete PCR reactions to amplify and detect target sequences of more than one virus, and (2) the lack of a rapid, nonhazardous means for specific det… Show more

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Cited by 18 publications
(3 citation statements)
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“…HIV DNA and Cell-Associated RNA Methods for quantification of HIV DNA and cell-associated RNA levels were previously published [21]; in brief, extracted PBMC DNA was measured using real-time PCR analysis targeting Gag DNA sequences; total amplifiable DNA (indicated cell input) was measured using real-time PCR analysis of a single-copy gene (the conserved region of the HLA-DQ alpha locus), as described elsewhere [29][30][31][32], and values were normalized per million PBMCs. Extracted cell-associated RNA levels were measured with the transcription-mediated amplification assay (Aptima; Gen-Probe), using modified PBMC extraction and transcription-mediated amplification of cell-associated HIV RNA [33,34].…”
Section: Gene Expression Profilingmentioning
confidence: 99%
“…HIV DNA and Cell-Associated RNA Methods for quantification of HIV DNA and cell-associated RNA levels were previously published [21]; in brief, extracted PBMC DNA was measured using real-time PCR analysis targeting Gag DNA sequences; total amplifiable DNA (indicated cell input) was measured using real-time PCR analysis of a single-copy gene (the conserved region of the HLA-DQ alpha locus), as described elsewhere [29][30][31][32], and values were normalized per million PBMCs. Extracted cell-associated RNA levels were measured with the transcription-mediated amplification assay (Aptima; Gen-Probe), using modified PBMC extraction and transcription-mediated amplification of cell-associated HIV RNA [33,34].…”
Section: Gene Expression Profilingmentioning
confidence: 99%
“…A number of studies have utilized multiplex PCRs for detection and differentiation of human retroviruses (45,46,99,111). Four primer pairs were combined to detect the gag region of HIV type 1 (HIV-1), the env region of HIV-2, the pol region of human T-cell leukemia virus type 1 (HTLV-1), and the tax region of HTLV-2 (45).…”
Section: Other Applicationsmentioning
confidence: 99%
“…The test was as sensitive as uniplex PCRs and allowed the detection of coinfection. Sunzeri et al (99) developed a multiplex PCR utilizing primer pairs targeting a portion of the gag region of HIV-1, the pol gene of HTLV-1 and -2, and a region of the HLA-DQ-␣ locus as an internal control. Products were analyzed by automated capillary DNA chromatography (products can also be separated and visualized using gel electrophoresis and ethidium bromide staining).…”
Section: Other Applicationsmentioning
confidence: 99%