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采用多重逆转录-PCR DNA 微阵列技术快速诊断中枢神经系统感染。

Rapid virological diagnosis of central nervous system infections by use of a multiplex reverse transcription-PCR DNA microarray.

机构信息

Clinical and Molecular Virology Unit, University Hospital, and EA-4303 Faculty of Medicine, Reims, France.

出版信息

J Clin Microbiol. 2011 Nov;49(11):3874-9. doi: 10.1128/JCM.01214-11. Epub 2011 Sep 14.

DOI:10.1128/JCM.01214-11
PMID:21918017
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3209095/
Abstract

Viruses are the main etiological cause of central nervous system (CNS) infections. A rapid molecular diagnosis is recommended to improve the therapeutic management of patients. The aim of this study was to evaluate the performances of a DNA microarray, the Clart Entherpex kit (Genomica, Coslada, Spain), allowing the rapid and simultaneous detection of 9 DNA and RNA neurotropic viruses: herpes simplex virus 1 (HSV-1), HSV-2, varicella-zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), human herpesvirus 6 (HHV-6), HHV-7, HHV-8, and the human enteroviruses (HEVs). This evaluation was performed with 28 samples from the European proficiency panels (Quality Control for Molecular Diagnostics [QCMD]; Glasgow, Scotland) and then with 78 cerebrospinal fluid (CSF) specimens. The majority of the QCMD results obtained by the DNA microarray were similar to those recorded by the overall QCMD participants. The main discrepant results were observed for low concentrations of HSV-2 and HEVs. From the clinical samples, the kit detected 27 of the 28 herpesvirus CNS infections and all of the 30 HEV-positive CSF samples. No false-positive result was observed among the 20 virus-negative CSF samples. The clinical sensitivity, specificity, and negative and positive predictive values of the assay were 98.3, 100, 95.2, and 100%, respectively, when the results were compared to those of commercially available PCR assays. Interestingly, HHV-7 was detected in 11 (37%) of the 30 HEV-positive CSF samples from children suffering from aseptic meningitis causing significantly longer lengths of stay at the hospital than infection with HEVs alone (2.4 versus 1.4 days; P = 0.038). In conclusion, this preliminary study showed that this DNA microarray could be a valuable molecular diagnostic tool for single and mixed DNA and RNA virus infections of the CNS.

摘要

病毒是中枢神经系统 (CNS) 感染的主要病因。建议进行快速分子诊断,以改善患者的治疗管理。本研究的目的是评估一种 DNA 微阵列(Clart Entherpex 试剂盒,Genomica,西班牙 Coslada)的性能,该微阵列允许快速且同时检测 9 种 DNA 和 RNA 神经嗜性病毒:单纯疱疹病毒 1 (HSV-1)、单纯疱疹病毒 2 (HSV-2)、水痘-带状疱疹病毒 (VZV)、巨细胞病毒 (CMV)、EB 病毒 (EBV)、人类疱疹病毒 6 (HHV-6)、HHV-7、HHV-8 和人类肠道病毒 (HEV)。这项评估是用来自欧洲能力验证小组(分子诊断质量控制 [QCMD];苏格兰格拉斯哥)的 28 个样本进行的,然后用 78 个脑脊液 (CSF) 标本进行。DNA 微阵列获得的大多数 QCMD 结果与总体 QCMD 参与者记录的结果相似。主要的差异结果观察到的是 HSV-2 和 HEV 的低浓度。从临床样本来看,该试剂盒检测到 28 例疱疹病毒 CNS 感染中的 27 例和 30 例 HEV 阳性 CSF 样本中的所有样本。在 20 例病毒阴性 CSF 样本中未观察到假阳性结果。与商业可用的 PCR 检测相比,该检测的临床灵敏度、特异性、阴性和阳性预测值分别为 98.3%、100%、95.2%和 100%。有趣的是,HHV-7 在 30 例 HEV 阳性 CSF 样本中检测到 11 例(37%),这些样本来自患有无菌性脑膜炎的儿童,与仅感染 HEV 的儿童相比,在医院的住院时间明显延长(2.4 天 vs. 1.4 天;P = 0.038)。总之,这项初步研究表明,这种 DNA 微阵列可能是一种有价值的分子诊断工具,可用于 CNS 的单一和混合 DNA 和 RNA 病毒感染。

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