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采用实时荧光定量 PCR 法对单纯疱疹病毒 1 型食管炎进行病毒学诊断。

Virological diagnosis of herpes simplex virus 1 esophagitis by quantitative real-time PCR assay.

机构信息

Department of Pathology, University Hospital of Reims, Reims, France.

出版信息

J Clin Microbiol. 2012 Mar;50(3):948-52. doi: 10.1128/JCM.05748-11. Epub 2011 Dec 14.

Abstract

Herpes simplex virus 1 (HSV-1) esophagitis diagnosis is routinely based on the endoscopic findings confirmed by histopathological examination of the esophagitis lesions. Virological diagnosis is not systematically performed and restricted to viral culture or to qualitative PCR assay from esophagitis biopsy specimens. The aim of this study was to assess the interest of quantitative real-time PCR assay in HSV-1 esophagitis diagnosis by comparing the results obtained to those of histological examination associated with immunohistochemical staining, which is considered the "gold standard." From 53 esophagitis biopsy specimens, the PCR assay detected HSV-1 in 18 of 19 histologically proven to have herpetic esophagitis and in 9 of 34 that had esophagitis related to other causes, demonstrating sensitivity, specificity, positive predictive value, and negative predictive value of 94.7%, 73%, 66.7%, and 96%, respectively. Interestingly, HSV-1 was not detected in 16 specimens without the histological aspect of esophagitis. The viral loads normalized per μg of total extracted DNA in each biopsy specimen detected positive by HSV PCR were then compared and appeared to be significantly higher in histopathologically positive herpetic esophagitis (median = 2.9 × 10(6) ± 1.1 × 10(8)) than in histopathologically negative herpetic esophagitis (median = 3.1 × 10(3) ± 6.2 × 10(3)) (P = 0.0009). Moreover, a receiver operating characteristics analysis revealed that a viral load threshold greater than 2.5 × 10(4) copies would allow an HSV-1 esophagitis diagnosis with a sensitivity and specificity of 83.3% and 100%, respectively. In conclusion, this work demonstrated that HSV quantitative PCR results for paraffin-embedded esophageal tissue was well correlated to histopathological findings for an HSV-1 esophagitis diagnosis and could be diagnostic through viral load assessment when histopathological results are missing or uncertain.

摘要

单纯疱疹病毒 1(HSV-1)食管炎的诊断通常基于内镜检查结果,并通过食管炎病变的组织病理学检查加以确认。病毒学诊断并未系统进行,仅限于病毒培养或对食管炎活检标本进行定性聚合酶链反应(PCR)检测。本研究旨在通过将定量实时 PCR 检测结果与组织学检查(联合免疫组织化学染色,这被认为是“金标准”)进行比较,评估其在 HSV-1 食管炎诊断中的应用价值。对 53 份食管炎活检标本进行检测,PCR 检测在 19 例经组织学证实为疱疹性食管炎的标本中检测到 HSV-1,在 34 例与其他原因相关的食管炎标本中检测到 9 例,其灵敏度、特异性、阳性预测值和阴性预测值分别为 94.7%、73%、66.7%和 96%。有趣的是,在没有食管炎组织学表现的 16 份标本中未检测到 HSV-1。对 HSV-PCR 检测为阳性的每份活检标本中总提取 DNA 进行病毒载量归一化,然后进行比较,在组织学阳性的疱疹性食管炎中明显更高(中位数=2.9×10^6±1.1×10^8),而在组织学阴性的疱疹性食管炎中(中位数=3.1×10^3±6.2×10^3)(P=0.0009)。此外,接受者操作特征分析显示,病毒载量阈值大于 2.5×10^4 拷贝数时,可使 HSV-1 食管炎诊断的灵敏度和特异性分别达到 83.3%和 100%。综上所述,本研究表明,石蜡包埋食管组织的 HSV 定量 PCR 结果与疱疹性食管炎的组织病理学发现具有良好的相关性,当组织学结果缺失或不确定时,可通过病毒载量评估进行诊断。

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