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No. 53 (PM 2)
HIV-1 RNA viral load in seminal fluid: quantification with NASBA
Lyana Setiawan, Agus Susanto Kosasih, Samsuridjal Djauzi, Haridana Indah Mahdi,
Runingsih
Dharmais Cancer Hospital, Indonesia
As the prevalence of HIV infection increases, the problem of discordant couples with HIV arises.
Although the number is yet unknown, discordant couple presented problem of possible infection to the
mothers and unborn babies. In this study, we assessed the utilization of nucleic acid sequence based
amplification (NASBA) to quantify HIV-1 RNA in seminal fluid, and reported the HIV-1 RNA viral
load in the seminal fluid of patients with discordant couple from the outpatient clinic of Dharmais
Cancer Hospital during July 2010 – February 2012, who have already been successfully treated with
antiretrovirals (ARV).
To determine the usefulness of NASBA, RNA from 2 unprocessed seminal fluid specimens were
extracted using the MiniMag®, amplified by NASBA, and the result quantified. To verify the usefulness
of sperm washing, we processed specimen with high HIV-1 RNA with gradient centrifugation to wash
the sperm, and quantified the viral load in the resulting seminal plasma from first centrifugation, and the
supernatant and sperm from the second washing. After optimization, we evaluated the result of 43
patients treated with ARV who were referred for HIV-RNA in seminal fluid test.
HIV-1 RNA was detected in the unprocessed seminal fluid (Pt#1 19,000 IU/ml; pt#2 740 IU/ml), and
in the seminal fluid from the first centrifugation. Supernatant and sperm from the second washing
showed HIV-RNA below detection limit. The result proved that boom extraction and NASBA principle
can be used for quantification of HIV-RNA in seminal fluid, and that sperm washing was effective to
minimize the risk of infection in assisted reproduction, but this needed to be verified with more samples.
All 43 patients showed HIV-RNA below detection limit (