Validating Sexual Behavior by Y-chromosome PCR

描述（由申请人提供）：估计报告偏倚和自我报告的有效性是性行为和安全套使用研究中的一个主要问题。作为生物标志物的偶发性性病是昂贵的，并且在低发病率人群中没有用。我们已经开发了一种高灵敏度的聚合酶链反应（PCR）检测精液Y染色体（Yc）序列在阴道液作为一个潜在的生物标志物的性行为和/或避孕套的效力。该检测试剂盒具有高度灵敏度，可在自我给药的阴道拭子上进行。试点数据发现，在无保护性交和禁欲期后，Yc可检测到15天，半衰期为3.4天。在本提案中，我们将通过在临床研究中招募50名女性来进一步表征该测定，以证明“概念证明”，即在避孕套保护的性交后，在性交后阴道中检测不到Yc。2）。为了确定不同条件下的检测灵敏度，我们建议进一步研究以评估阴道微环境的特征，这可能影响Yc清除。我们假设月经和冲洗会缩短Yc清除时间。首先，我们将通过在120名女性中仔细监测的性交后2周内评估Yc清除率来评估月经的存在/不存在对Yc清除率的影响，这些女性将作为自己的对照参与-她们将参与两次（总共224个周期），比较在测量间隔Yc清除时间内的月经期与没有月经的时间间隔的影响。我们将通过比较120名妇女在冲洗后的Yc清除率，与同一受试者在未冲洗的重复研究间隔（共224个研究周期）相比，评价冲洗的效果，冲洗是巴尔的摩有STD风险的妇女的常见做法。从我们1990- 1992年的避孕套使用研究中，我们收集了82名在2周内报告性行为且100%使用避孕套的妇女和322名未使用避孕套的妇女的宫颈阴道灌洗液。将对这些样本进行Yc分析，并与人口统计学和行为学数据以及访视时宫颈炎症和STD的伴随标志物进行比较。结果将与先前发表的研究结果进行比较，该研究使用STD发病率作为生物标志物变量。由于Yc采样仅需要阴道拭子，并且在现场条件下稳定，因此该测试有望成为性行为研究的实用且易于使用的生物标志物。

DESCRIPTION (provided by applicant): Estimating reporting bias and validity of self-reports are a major issue in sexual behavior and condom use research. Incident STDs as biomarkers are expensive, and not useful in low-incident populations. We have developed a highly sensitive polymerase chain reaction (PCR) assay for semen Y-chromosome (Yc) sequences in vaginal fluid as a potential biomarker of sexual behavior and/or condom efficacy. The assay is highly sensitive and can be performed on a self-administered vaginal swab. Pilot data found that after unprotected intercourse followed by an abstinence period, Yc was detectable for 15 days, and the half-life was 3.4 days. In this proposal, we will further characterize the assay by enrolling 50 women in a clinical study to demonstrate "proof of concept", i.e. that after condom-protected intercourse no Yc is detectable in the post-coital vagina. 2). To define assay sensitivity under different conditions, we propose further research to evaluate characteristics of the vaginal microenvironment, which could affect Yc clearance. We hypothesize that menses and douching would shorten Yc clearance times. First we will evaluate the presence/absence of menses on Yc clearance by evaluating Yc clearance over a carefully monitored 2 week post-coital period in 120 women who will participate as their own control-they will participate twice (for a total of 224 cycles), comparing the effect of having their menstrual period during the measurement interval Yc clearance time compared to a time interval where menses did not occur. We will evaluate the effect of douching, a common practice in women in Baltimore at risk for STD, by comparing Yc clearance in 120 women after douching, compared to a repeated study interval in the same subject where they did not douche (a total of 224 study cycles). From our 1990-92 studies of condom use, we have archived cervicovaginal lavage fluid from 82 women reporting sex within 2 weeks and 100% condom use, and 322 who did not use condoms. These samples will be assayed for Yc and compared with demographic and behavioral data, and concomitant markers of cervical inflammation and STDs at time of visit. The outcomes will be compared to the findings from a previously published study that used STD incidence as the biomarker variable. Since Yc sampling requires only a vaginal swab, and is stable under field conditions, this test promises to be a practical and easily used biomarker for sexual behavior research.