An evaluation of semen processing methods for eliminating HIV-1

Objective: Whether artificial insemination can provide adequate protection for discordant couples where the man is HIV-1 positive and the woman is HIV -1 negative is uncertain because of the paucity of HIV-1 elimination data a ssessing current sperm-washing techniques. We evaluated how effectively the se techniques eliminate HIV-1 RNA and proviral DNA from semen. Methods: Spermatozoa were separated from semen samples from HIV-1-positive patients with haemophilia by discontinuous Percoll gradient centrifugation and the 'swim-up' method. The HIV-1 RNA and proviral DNA were measured by a highly sensitive PCR. In another test 5 x 10(6) copies of HIV-1 RNA (LAI s train) were added to semen from healthy donors and then assessed after sing le and combined procedures. Results: Swim-up processing after Percoll gradient centrifugation reduced H IV-1 RNA and HIV-1 proviral DNA in semen to undetectable levels in the orig inal specimen. Although discontinuous and continuous Percoll gradient centr ifugation respectively reduced HIV-1 RNA added to seminal plasma specimens from healthy donors to less than < 1 copy from 10(5) and about 1 copy per 1 0(3) pre-separation copies, the discontinuous method left detectable HIV-1 RNA and proviral DNA in one out of 12 samples from patients with HIV-1 infe ction (8%). HIV-1 RNA and proviral DNA were decreased to undetectable level s after adding the swim-up procedure. Conclusions: Swim-up separation following Percoll gradient centrifugation s hould offer adequate protection for HIV-1-discordant couples. (C) 2000 Lipp incott Williams & Wilkins.