Hobson A, Wald A, Wright N, Corey L
Department of Laboratory Medicine, University of Washington, Seattle, USA.
J Clin Microbiol. 1997 Mar;35(3):548-52. doi: 10.1128/jcm.35.3.548-552.1997.
Previous studies have shown an association between the approximate titer of herpes simplex virus (HSV) DNA in clinical specimens and the ability to isolate HSV from genital secretions. To control for variance in amplification conditions, we developed a competitive quantitative PCR (QC PCR) for the detection of HSV DNA. The assay accurately measured from 10 to 10(6) copies of HSV DNA. We compared the QC PCR with our previous semiquantitative detection method and found concordance for 61 of 63 positive specimens. We also evaluated the HSV DNA content from individual swabs of genital secretions obtained from individual sites of the genital tract (cervix, vulva, and rectum) with that from one swab with secretions from all three sites. The concordance for detecting HSV DNA was 91%; for only 4 of 143 collection days was there a > 1 log difference between the two collection methods. A single swab with secretions from all three genital sites and evaluated in a QC PCR format can accurately measure the frequency of subclinical and clinical shedding of HSV and the titer of HSV shed from the genital region. Such an approach should be very useful in the evaluation of antiviral chemotherapy for HSV.
先前的研究表明，临床标本中单纯疱疹病毒（HSV）DNA的大致滴度与从生殖器分泌物中分离出HSV的能力之间存在关联。为了控制扩增条件的差异，我们开发了一种用于检测HSV DNA的竞争性定量PCR（QC PCR）。该检测方法能准确测量10至10⁶拷贝的HSV DNA。我们将QC PCR与我们先前的半定量检测方法进行了比较，发现63份阳性标本中有61份结果一致。我们还评估了从生殖道各个部位（宫颈、外阴和直肠）采集的单个生殖器分泌物拭子中的HSV DNA含量，并与采集所有三个部位分泌物的一个拭子中的含量进行了比较。检测HSV DNA的一致性为91%；在143个采集日中，只有4天两种采集方法之间的差异>1个对数。用QC PCR方法评估采集所有三个生殖器部位分泌物的单个拭子，能够准确测量HSV亚临床和临床排毒的频率以及从生殖器区域排出的HSV滴度。这种方法在评估HSV抗病毒化疗方面应该非常有用。
