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 assessing current sperm-washing techniques. We evaluated how effectively these 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 strain) were added to semen from healthy donors and then assessed after single and combined procedures.


RESULTS
Swim-up processing after Percoll gradient centrifugation reduced HIV-1 RNA and HIV-1 proviral DNA in semen to undetectable levels in the original specimen. Although discontinuous and continuous Percoll gradient centrifugation 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 10(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 infection (8%). HIV-1 RNA and proviral DNA were decreased to undetectable levels after adding the swim-up procedure.


CONCLUSIONS
Swim-up separation following Percoll gradient centrifugation should offer adequate protection for HIV-1-discordant couples.
Hanabusa
H
H
Department of Hematology, Ogikubo Hospital, Tokyo, Japan.
Kuji
N
N
Kato
S
S
Tagami
H
H
Kaneko
S
S
Tanaka
H
H
Yoshimura
Y
Y
D000328:Adult; D002469:Cell Separation; D002499:Centrifugation, Density Gradient / Q000379:methods; D004279:DNA, Viral / Q000032:analysis*; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D011205:Povidone; D012367:RNA, Viral / Q000032:analysis*; D012661:Semen / Q000821:virology*; D012822:Silicon Dioxide; D013094:Spermatozoa / Q000821:virology*
D004279:DNA, Viral; D012367:RNA, Viral; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone