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Detection of human immunodeficiency virus type 1 by using the polymerase chain reaction and a time-resolved fluorescence-based hybridization assay. Dahlén PO, Iitiä AJ, Skagius G, Frostell A, Nunn MF, Kwiatkowski M.Detection of HIV-1 DNA in crude cell lysates of peripheral blood mononuclear cells by the polymerase chain reaction and nonradioactive oligonucleotide probes. Conway B, Adler KE, Bechtel LJ, Kaplan JC, Hirsch MS.Decrease in HIV provirus in peripheral blood mononuclear cells during zidovudine and human rIL-2 administration. Clark AG, Holodniy M, Schwartz DH, Katzenstein DA, Merigan TC.Detection and typing of human papillomaviruses in cervical smears by an original application of the polymerase chain reaction. Charlotte F, Olivier JL, Chypre C, Beuret T, Sadoul G, Chatelet F, Luboinski J, Marchand J, Bereziat G.The Italian Study Group on Occupational Risk of HIV Infection. Zidovudine prophylaxis after accidental exposure to HIV: the Italian experience.
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We also assessed the usefulness of this test for the estimation of the HIV-1 DNA load by the end point dilution method with serial dilutions of blood lysates from 26 HIV-1-seropositive patients. The results obtained in 54 seropositive and 120 seronegative individuals demonstrated the ability of the AMPLICIS test to be used for HIV-1 provirus detection: 53 of 54 of the seropositive specimens were found to be positive with at least two primer sets. Three new primer sets located in the gag and pol structural genes and in the tat regulatory gene of HIV-1 were studied. The solid-phase capture and counting of the final amplified products allowed us to define precise interpretive criteria to determine the positivity level of the test. In order to simplify the general process, whole-blood lysates rather than peripheral blood mononuclear cell lysates were used for the amplifications. To identify the HIV-1 proviral DNA in blood, we used a solid-phase assay based on the affinity capture and the gamma counting of the amplified product after a nested polymerase chain reaction (AMPLICIS test). The routine detection of human immunodeficiency virus type 1 (HIV-1) proviral DNA in clinical samples requires a standardized, simple, and sensitive test.
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