Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, 14115-111, Tehran, Iran.
J Virol Methods. 2012 Aug;183(2):170-5. doi: 10.1016/j.jviromet.2012.04.010. Epub 2012 May 4.
Quantitation of human cytomegalovirus (HCMV) DNA is used to monitor immunocompromised patients in order to identify patients for preemptive therapy. Although several commercial qPCR assays are available for quantitation of HCMV, their major disadvantage is the high cost. In the present study, an internally controlled quantitative real-time PCR assay based on hydrolysis probe technology was developed for detection and quantitation of HCMV DNA in plasma samples. To demonstrate its performance characteristics, a total of 178 plasma samples from 102 kidney and hematopoietic stem cell transplanted patients were tested. The assay showed good precision and reproducibility, and an analytical sensitivity of 288.5 copies/ml or 17.6 copies/reaction. A sensitivity of 93.1% and a specificity of 96.6% were determined by examining clinical samples. Analysis of a panel containing potentially interfering viruses demonstrated no cross-reactivity with the assay. A strong correlation was observed between this qPCR method and the commercial Artus(®) CMV RG PCR kit (R=0.948; P=0.000). These results indicate that the affordable internally controlled qPCR method described will be useful for monitoring HCMV infection in plasma samples of immunocompromised patients.
人巨细胞病毒（HCMV）DNA 的定量检测用于监测免疫功能低下的患者，以确定需要进行抢先治疗的患者。虽然有几种商业化的 qPCR 检测方法可用于 HCMV 的定量检测，但它们的主要缺点是成本高。在本研究中，开发了一种基于水解探针技术的内部对照定量实时 PCR 检测方法，用于检测和定量血浆样本中的 HCMV DNA。为了证明其性能特点，共检测了 102 例肾和造血干细胞移植患者的 178 份血浆样本。该检测方法具有良好的精密度和重现性，分析灵敏度为 288.5 拷贝/ml 或 17.6 拷贝/反应。通过检测临床样本确定了 93.1%的灵敏度和 96.6%的特异性。分析包含潜在干扰病毒的面板表明该检测方法与该检测无交叉反应。该 qPCR 方法与商业化的 Artus(®)CMV RG PCR 试剂盒之间存在很强的相关性（R=0.948；P=0.000）。这些结果表明，所描述的这种经济实惠的内部对照 qPCR 方法将有助于监测免疫功能低下患者的血浆样本中的 HCMV 感染。
