HIV/AIDS Assisted Reproduction: PCR detects low HIV-1 RNA in semen preparations used for assisted reproduction

AIDSWEEKLY Plus; Monday, September 18, 2006
Staff Medical Writers

In a recently published report, investigators in Belgium conducted laboratory-based research in a university hospital to "develop a method for same-day validation of processed, semen in the setting of assisted reproductive techniques (ART) with patients who are seropositive for human immunodeficiency virus, type 1 (HIV-1)."

Study subjects included HIV-1 seronegative and seropositive volunteers. The study included evaluation "of the sensitivity of a reverse-transcriptase (RT)-nested polymerase chain reaction (PCR) in HIV-1 RNA-positive blood plasma, in artificially infected blood plasma and semen; and in 85 semen samples of 29 HIV-1-seropositive volunteers.

Semen was submitted to gradient separation, followed by swim-up. Qualitative detection of HIV-1 RNA in blood plasma and in different parts of semen preparation by using RT-nested PCR, PCR inhibition control by dilution of samples, and an internal control," said the authors.

"The detection limit of our PCR was 20 HIV-1 RNA copies per milliliter. Among seropositive patients RNA was detected in 25% of fresh semen, 36.5 of seminal plasma, 27.5% of gradient supernatants, and 7.1% of final preparations before the migration-sedimentation stage.

"Positive final, preparations were observed in patients who had blood viral loads of greater than or equal to20,000 HIV-1 RNA copies per milliliter. Inhibition was present in 17.6% of seminal plasma and in 20% gradient supernatants and in 2 final preparation's among 69 tested," reported B. Lesage and colleagues at the Free University of Brussels.

"Among preparations tested after the migration sedimentation stage," continued scientists, "2 were positive (1 patient; 70,000 HIV-1 RNA copies per milliliter)."

"The RT-nested PCR detects low viral load and allows the validation of semen preparations of HIV-1-seropositive patients for ART on the day of sampling. For this purpose, the validation is performed on spermatozoa that are obtained, after gradient separation before swim-up.

"Inhibition of the PCR must be controlled by using an internal control that is well-designed to explore the detection limit of the method," Lesage concluded.

Lesage and colleagues published the results of their research in Fertility and Sterility (Development and evaluation of a qualitative reverse-transcriptase nested polymerase chain reaction protocol for same-day viral validation of human immunodeficiency virus type 1 ribonucleic acid in processed semen. Fertil Steril. 2006 Jul;86(1):121-8).

For additional information, contact Y. Englert, Free University of Brussels, Hopital Erasme, 808 Route Lennik, B-1070 Brussels, Belgium.

The publisher of the journal Fertility and Sterility can be contacted at: Elsevier Science Inc., 360 Park Avenue South, New York, NY 10010-1710, USA.

Keywords: Brussels, Belgium, HIV/AIDS, Nested PCR, Assisted Reproduction, Viral Load, Assay Validation.

This article was prepared by AIDS Weekly editors from staff and other reports.

Reference

Lesage B, Vannin AS, Emiliani S, et al., “Development and evaluation of a qualitative reverse-transcriptase nested polymerase chain reaction protocol for same-day viral validation of human immunodeficiency virus type 1 ribonucleic acid in processed semen”, Fertil Steril. 2006 Jul;86(1):121-8.

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