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Human immunodeficiency virus load. Quantitative assessment in semen from seropositive individuals and in spiked
1Department of Pathology, University of Southern California School of Medicine, Los Angeles 90032-3626, USA.
Insights
Quantifying human immunodeficiency virus (HIV) in semen is crucial. Reverse transcription polymerase chain reaction (RT-PCR) offers a highly sensitive method for detecting HIV RNA in seminal plasma, cells, and whole semen.
Area of Science:
- Virology
- Reproductive Health
- Molecular Diagnostics
Background:
- Accurate quantification of human immunodeficiency virus (HIV) in semen is essential for understanding transmission dynamics and evaluating treatment efficacy.
- Existing methods for HIV detection in seminal samples have limitations in sensitivity and scope.
Purpose of the Study:
- To establish reliable criteria for quantifying HIV in seminal plasma, seminal cells, and whole semen.
- To standardize and compare the sensitivity of reverse transcription polymerase chain reaction (RT-PCR), DNA-PCR, and semen HIV culture assays.
Main Methods:
- Standardization of RT-PCR, DNA-PCR, and semen HIV culture assays using spiked seminal plasma with known HIV titers.
- Assessment of viral quantity in freshly collected seminal cells and seminal plasma from HIV-infected individuals.
Main Results:
- HIV was isolated from 76% of peripheral blood mononuclear cells (PBMCs) and 24% of ejaculates from HIV-seropositive men.
- RT-PCR demonstrated 5-125 times greater sensitivity than cell cultures for HIV quantitation in various seminal fluid components.
- HIV-RNA was detected in samples with higher viral dilutions than those detectable by culture.
Conclusions:
- Cell-free HIV is present in seminal fluid in quantities exceeding culturable virus.
- High-sensitivity HIV-RNA detection using RT-PCR is effective for direct viral quantitation in seminal plasma, 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 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.