S Rasheed1, Z Li, D Xu. 1. Department of Pathology, University of Southern California School of Medicine, Los Angeles 90032-3626, USA.
Abstract
OBJECTIVE: To establish criteria for the quantitation of the human immunodeficiency virus (HIV) in seminal plasma, seminal cells and the whole semen of HIV-infected individuals. The reverse transcription polymerase chain reaction (RT-PCR), DNA-PCR and semen HIV culture assays were standardized by testing seminal plasma spiked separately with serial dilutions of cell-free and cell-associated HIV stocks of known titers. The standardized assays were then used to assess the quantity of virus in the freshly collected seminal cells and seminal plasma. RESULTS: Analysis of freshly collected peripheral blood mononuclear cells (PBMCs) and paired semen from HIV-seropositive men who had received antiviral drugs and/or immunemodulators indicated that HIV could be isolated from 42 of 55 (76%) samples of peripheral blood mononuclear cells (PBMCs) and 13 of 55 (24%) samples of ejaculates. Since no semen sample was culture positive in the absence of culturable HIV in PBMCs of the same individual, RT-PCR was 5-125 times more sensitive than cell cultures for the quantitation of HIV spiked in seminal plasma, freshly collected seminal fluid and whole semen. Further, HIV-RNA was detected in samples containing higher dilutions of virus from which HIV was not isolated by culture. CONCLUSION: We conclude that cell-free HIV is present in excess of the culturable virus in all specimens tested and that the high sensitivity of HIV-RNA detection is useful for quantitation of the virus directly in seminal fluid, seminal cells and whole semen.
OBJECTIVE: To establish criteria for the quantitation of the human immunodeficiency virus (HIV) in seminal plasma, seminal cells and the whole semen of HIV-infected individuals. The reverse transcription polymerase chain reaction (RT-PCR), DNA-PCR and semen HIV culture assays were standardized by testing seminal plasma spiked separately with serial dilutions of cell-free and cell-associated HIVstocks of known titers. The standardized assays were then used to assess the quantity of virus in the freshly collected seminal cells and seminal plasma. RESULTS: Analysis of freshly collected peripheral blood mononuclear cells (PBMCs) and paired semen from HIV-seropositivemen who had received antiviral drugs and/or immunemodulators indicated that HIV could be isolated from 42 of 55 (76%) samples of peripheral blood mononuclear cells (PBMCs) and 13 of 55 (24%) samples of ejaculates. Since no semen sample was culture positive in the absence of culturable HIV in PBMCs of the same individual, RT-PCR was 5-125 times more sensitive than cell cultures for the quantitation of HIV spiked in seminal plasma, freshly collected seminal fluid and whole semen. Further, HIV-RNA was detected in samples containing higher dilutions of virus from which HIV was not isolated by culture. CONCLUSION: We conclude that cell-free HIV is present in excess of the culturable virus in all specimens tested and that the high sensitivity of HIV-RNA detection is useful for quantitation of the virus directly in seminal fluid, seminal cells and whole semen.
Authors: S A Fiscus; D Brambilla; R W Coombs; B Yen-Lieberman; J Bremer; A Kovacs; S Rasheed; M Vahey; T Schutzbank; P S Reichelderfer Journal: J Clin Microbiol Date: 2000-06 Impact factor: 5.948
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Authors: Ruizhong Shen; Ernesto R Drelichman; Diane Bimczok; Christina Ochsenbauer; John C Kappes; Jamie A Cannon; Daniela Tudor; Morgane Bomsel; Lesley E Smythies; Phillip D Smith Journal: J Immunol Date: 2010-03-05 Impact factor: 5.422