Marshall D S, Linfert D R, Draghi A, McCarter Y S, Tsongalis G J
Department of Pathology and Laboratory Medicine, Hartford Hospital, Hartford, Connecticut 06102, USA.
Mod Pathol. 2001 Mar;14(3):152-6. doi: 10.1038/modpathol.3880273.
Genital herpes simplex virus (HSV) is of major public health importance, as indicated by the marked increase in the prevalence of genital herpes over the past two decades. Viral culture has traditionally been regarded as the gold standard for diagnosis. In this study, we compared viral culture and the amplification of HSV DNA by the polymerase chain reaction (PCR) with respect to sensitivity, cost, clinical utility, and turnaround time. Patient sample swabs from 100 individuals were inoculated onto MRC-5 cells for isolation. Positive results were confirmed via a direct fluorescent antibody technique, and serotyping, when requested, was performed using HSV-1 and -2-type-specific sera. PCR techniques employed an extraction step of the same initial swab specimen, followed by PCR amplification, using a multiplex assay for HSV-1, 2 DNA. HSV-positive results were found in 32/100 samples via culture and in 36/100 samples via PCR. PCR-positive results yielded 16 (44%) patients infected with HSV-1 and 20 (56%) patients infected with HSV-2. Turnaround time for viral culture averaged 108 hours for positive results and 154 hours for negative results; PCR turnaround time averaged 24--48 hours. Laboratory cost using viral culture was $3.22 for a negative result and $6.49 for a positive result (including direct fluorescent antibody). Serotyping added $10.88 to each culture-positive test. Although laboratory costs for PCR were higher at $8.20/sample, reimbursement levels were also higher. We propose a multiplex PCR assay for diagnosis of HSV-1 and HSV-2 from patient swabs for use in a routine clinical laboratory setting. This assay offers increased sensitivity, typing, and improved turnaround time when compared with traditional viral culture techniques. Although it appears that PCR testing in a routine clinical laboratory setting is cost prohibitive compared with the case of nonserotyped viral culture, it may be very useful when clinical utility warrants distinguishing between HSV 1 and 2 and may be cost effective when reimbursement issues are examined.
生殖器单纯疱疹病毒（HSV）具有重大的公共卫生意义，过去二十年中生殖器疱疹患病率的显著上升就表明了这一点。传统上，病毒培养一直被视为诊断的金标准。在本研究中，我们比较了病毒培养和聚合酶链反应（PCR）扩增HSV DNA在敏感性、成本、临床实用性和周转时间方面的差异。将100名患者的样本拭子接种到MRC - 5细胞上进行分离。通过直接荧光抗体技术确认阳性结果，如有需要，使用HSV - 1和 - 2型特异性血清进行血清分型。PCR技术采用相同初始拭子标本的提取步骤，然后进行PCR扩增，使用针对HSV - 1、2 DNA的多重检测方法。通过培养在32/100的样本中发现HSV阳性结果，通过PCR在36/100的样本中发现阳性结果。PCR阳性结果显示，16名（44%）患者感染HSV - 1，20名（56%）患者感染HSV - 2。病毒培养阳性结果的平均周转时间为108小时，阴性结果为154小时；PCR周转时间平均为24 - 48小时。病毒培养的实验室成本，阴性结果为3.22美元，阳性结果为6.49美元（包括直接荧光抗体）。血清分型给每个培养阳性检测增加10.88美元。虽然PCR的实验室成本更高，为8.20美元/样本，但报销水平也更高。我们提出一种用于从患者拭子中诊断HSV - 1和HSV - 2的多重PCR检测方法，用于常规临床实验室环境。与传统病毒培养技术相比，该检测方法具有更高的敏感性、可分型性，并缩短了周转时间。尽管在常规临床实验室环境中，PCR检测似乎比未进行血清分型的病毒培养成本更高，但当临床实用性需要区分HSV 1和2时，它可能非常有用，并且在考虑报销问题时可能具有成本效益。
