Direct detection of infectious HIV-1 in blood using a centrifugation-indicator cell assay.
Plasma HIV RNA is a useful surrogate marker for predicting HIV-1 disease progression in infected individuals but provides no information regarding the infectious viral titer. Traditional assays of infectious HIV-1 are, however, time consuming, insensitive, use non-standardized reagents and are subject to selection bias introduced by prolonged cultivation. In this pilot study infectious HIV-1 was detected directly in patient plasma using the indicator line HeLa-CD4-CCR5-LTR/beta-gal in a centrifugation-culture method. Replication competent HIV-1 was identified within 2 days of tissue culture inoculation in six (26%) of 23 plasma specimens. The capability of a new cell line, MT4-CCR5-tat, to amplify plasma HIV-1 was also tested. HIV was cultivated from ten (71%) of 14 specimens using MT4-CCR5-tat cells before titering the virus with the indicator cell assay. Using these stable cell lines in refined versions of this assay it may be feasible to develop rapid, simple methods for titering infectious plasma HIV-1 and for testing the susceptibility of the virus to antiretroviral drugs.
Department of Medicine, University of Washington School of Medicine, Seattle, USA. rdh@u.washington.edu
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Harrington
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Wu
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Pullen
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Emerman
D001769:Blood / Q000821:virology*; D015704:CD4 Antigens / Q000378:metabolism; D002460:Cell Line; D002498:Centrifugation; D015658:HIV Infections / Q000821:virology*; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D010865:Pilot Projects; D012367:RNA, Viral / Q000097:blood; D019713:Receptors, CCR5 / Q000378:metabolism; D014766:Viremia / Q000821:virology; D014776:Virus Cultivation; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D015704:CD4 Antigens; D012367:RNA, Viral; D019713:Receptors, CCR5; D001616:beta-Galactosidase