Validating Sexual Behavior by Y-chromosome PCR

描述（由申请人提供）：评估自我报告的报告偏差和有效性是性行为和避孕套使用研究中的一个主要问题。偶发性传播疾病作为生物标志物是昂贵的，而且在低发病率人群中没有用处。我们开发了一种高度敏感的聚合酶链反应（PCR）检测阴道液中精液y染色体（Yc）序列，作为性行为和/或避孕套有效性的潜在生物标志物。该分析是高度敏感的，可以在自己给药的阴道拭子上进行。试点数据发现，在无保护性交后，禁欲期，Yc可检测15天，半衰期为3.4天。在本提案中，我们将通过招募50名妇女进行临床研究来进一步表征该检测方法，以证明“概念证明”，即在有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可能影响Yc清除的阴道微环境特征。我们假设月经和冲洗会缩短Yc清除时间。首先，我们将评估月经是否存在对Yc清除率的影响，方法是对120名女性进行精心监测的2周后的Yc清除率评估，这些女性将作为自己的对照——她们将参加两次（总共224个周期），比较在测量间隔Yc清除率期间月经的影响，与未发生月经的时间间隔的影响。我们将通过比较120名女性在冲洗后的Yc清除率来评估冲洗的效果，这是巴尔的摩有性病风险的女性的一种常见做法，与同一受试者中不冲洗的重复研究间隔（总共224个研究周期）进行比较。从我们1990-92年关于避孕套使用的研究中，我们收集了82名妇女的宫颈阴道灌洗液，这些妇女报告在2周内发生性行为，100%使用避孕套，322名没有使用避孕套。将对这些样本进行Yc检测，并与人口统计学和行为数据以及访问时宫颈炎症和性传播疾病的伴随标志物进行比较。这些结果将与先前发表的一项使用性病发病率作为生物标志物变量的研究结果进行比较。由于Yc取样只需要阴道拭子，并且在现场条件下稳定，因此该测试有望成为一种实用且易于使用的性行为研究生物标志物。

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.