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Try out PMC Labs and tell us what you think. Learn More. Genitourinary tract samples are required to investigate male HIV-1 infectivity. HIVseropositive men were evaluated after 48 hours of sexual abstinence. Participants provided semen specimens one week later. An audio computer-assisted self-interview ACASI administered after each specimen collection evaluated acceptability, adherence to instructions, and recent genitourinary symptoms. Concordance of at repeat visits was Sex partners of seropositive men acquire HIV-1 infection via direct contact with semen. Therefore, alternative sampling methods are needed to define the determinants of male HIV-1 shedding and to improve strategies to reduce transmission.

Optimal methods to sample the male genital tract remain unclear, especially for prospective studies requiring regular specimen collections. HIVseropositive Kenyan men were recruited from a high-risk cohort. Blood was collected for CD4 count and plasma viral load testing. A standardized examination was performed, including urethral swab collection. After the examination, a male or female clinician performed prostatic massage until expressed prostatic secretions EPS were visualized at the urethral meatus or for up to 5 minutes.

As recommended by Price et al , 9 participants could provide semen in clinic or at home. Participants were instructed to submit specimens to the laboratory within 2 hours of ejaculation. Participants completed an audio computer-assisted self-interview ACASI in Kiswahili after each sample collection to evaluate acceptability, comfort, adherence to the prescribed abstinence period, and genitourinary symptoms in the past week. Supernatant aliquots were transferred into labelled 1.

Semen was processed immediately after submission. Volume, appearance and degree of liquefaction were noted, and the specimen was warmed in an incubator until complete liquefaction before processing. A wet-mount was examined under 40x power to estimate sperm motility. Aliquots were diluted in distilled water for haemocytometer sperm and round cell counts. Laboratory procedures were developed by a qualified andrologist CHM , and specimen cell-counting quality assured by the University of Washington Male Fertility Laboratory. If two complete sample sets were available for a participant, both sets were tested to evaluate stability of over time.

Factors associated with success of each sample collection type were evaluated by logistic regression using generalized estimating equations GEE with an exchangeable correlation matrix. Values below the quantification limit were set to half the lower limit for that specimen type.

To assess within-subject variability, the standard deviation of the difference in log 10 HIV-1 RNA was calculated across repeated visits for each sample type; this analysis was restricted to samples with detectable HIV-1 RNA at one or both visits.

Analyses employed Stata version Participants gave informed consent, following procedures approved by committees of the Kenya Medical Research Institute and University of Washington. Between November and July , 52 men enrolled Table 1. Participants were older than the 43 men who attended clinic during the study period but did not enroll median age Participants were less likely to be single, although this difference was borderline There were no differences in education, sexual orientation, ethnicity, or religion between groups.

Participants attended quarterly visits until July 31, , completion of five visits, or enrolment in a sub-study of men initiating ART. Forty-seven men had visits contributing to the present analysis; the other five participated only in the sub-study. Participants attended a median of 4 visits inter-quartile range [IQR], 2 — 5 visits. Overall, of Nine participants Reported adherence to the prescribed abstinence period was For semen collections, median acceptability was 10 IQR 10 — 10, range 0 — 10 , with reservations reported at 6.

For semen collections, median discomfort was 0 IQR 0 — 0, range 0 — 7 , with some discomfort reported at 3. Difficulty producing semen was reported at two collections 1. Reasons documented for 13 missed semen collections Semen was collected after of visits Most semen samples were provided in clinic Both specimens were collected at least once from 35 men One man 2.

Success was less likely with the female clinician and for repeat visits. Transactional sex was collinear with sex with men, and is omitted. Ten participants were taking ART, including nine who received a standard first-line regimen of stavudine, lamivudine, and efavirenz and one who received a second-line regimen of tenofovir, abacavir, and boosted lopinavir. Additional sample sets were collected at repeat visits from 19 men, for a total of 48 paired sample sets. Concordance of test at 19 repeat visits was At the initial visit, 10 of 29 men Thirteen men were similar 1.

Most participants rated discomfort as low for both sampling methods. Success was less likely on repeat visits or when the clinician was female. Despite initial concerns that Kenyan men would not find semen collection by masturbation acceptable, successful semen collection in this study compared favorably with a study of semen collection in Malawi, in which of men In contrast, our population included mostly MSM and transactional sex workers, with lower CD4 count proving the only factor related to unsuccessful semen collection.

Decreased semen quality has been noted among HIVseropositive men with advanced immunosuppression. Our study likely underestimates the extent of this problem, as men with a history of ejaculatory difficulties are unlikely to enroll in studies requiring provision of semen samples. Blood plasma HIV-1 RNA levels were relatively low even in untreated men, who were healthy volunteers able to provide semen. When men with very low genital HIV-1 levels are studied, are influenced by the HIV-1 RNA quantification limits in semen as compared to other fluids such as blood or urine.

Training of clinicians was completed in one day, and both clinicians felt comfortable with the procedure after performing massage on two or three volunteer patients. Therefore, for certain men and in certain circumstances e.

We do not know if seminal HIV-1 RNA level represents an independent predictor of transmission at a given blood plasma viral load. Other limitations of this study include limited sample size, uniqueness of the population studied, incomplete ACASI data, and lack of randomization of collection method order, which might have caused minor differences in HIV-1 RNA levels, particularly if adherence to the prescribed abstinence period differed. This observation supports the hypothesis that most HIV-1 replication occurs in periurethral tissue rather than in glandular structures contributing to the ejaculate.

We are indebted to the men who participated in this study. The contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH. Competing interests. None declared for all authors. Licence agreement. National Center for Biotechnology Information , U. Sex Transm Infect. Author manuscript; available in PMC Apr 1. Susan M.

Krieger , MD, 1 Peter L. Muller , PhD, 1 Sarah E. Holte , PhD, 1, 5 Kishor N. John N. Peter L. Lorraine W. Kelly M. Joan A. Charles H. Sarah E. Kishor N. Scott McClelland. Norbert M. Eduard J. Robert W. Author information Copyright and information Disclaimer. Corresponding author and request for reprints: Susan M. Copyright notice. The publisher's final edited version of this article is available at Sex Transm Infect. See other articles in PMC that cite the published article.

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