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用于检测临床样本中BK病毒和JC病毒的内部实时荧光定量PCR方法与商业实时荧光定量PCR方法的比较评估

Comparative Evaluation of In-House and Commercial Real-Time PCR Methods for the Detection of the BK and JC Viruses in Clinical Samples.

作者信息

Özdemir Mehmet, Ayan Uğur, Şevik Murat

机构信息

Department of Medical Microbiology, Division of Medical Virology, Meram Medical Faculty, Necmettin Erbakan University, Konya, Turkey.

Medical Microbiology Laboratory, Istanbul Medeniyet University, Göztepe Training and Research Hospital, Istanbul, Turkey.

出版信息

J Lab Physicians. 2020 Aug;12(2):79-83. doi: 10.1055/s-0040-1716603. Epub 2020 Sep 2.

Abstract

The two most common human polyomaviruses are the BK (BKV) and JC viruses (JCV). Diseases associated with polyomavirus usually occur in cases of severe cellular immunosuppression. BKV and JCV can cause many diseases, especially if they are reactivated in an immunosuppressed host. The aim of this study is to compare and evaluate the results of real-time polymerase chain reaction (PCR) methods targeting the small and large T gene regions of the viral genome, considering polymorphisms occurring in the viral genome of BKV and JCV.  Urinary specimens of 82 patients were taken from immunosuppressed patient and sent to molecular microbiology laboratory of Meram Medical Faculty. The small t gene was investigated using a commercial kit (LightMix, Roche) by real-time PCR method. Large T gene was investigated by using the optimized in-house real-time PCR method. Sequence analysis was accepted as the standard method.  BKV positivity was detected in 9 samples and JCV positivity in 61 samples by real-time PCR method specific to small t gene region; BKV positivity in 21 samples and JCV positivity in 67 samples were determined by real-time PCR method specific to the large T gene region. Statistically, there was a significant difference for BKV, but not significant difference for JCV detection between the two methods.  Different polymorphisms in the target gene regions were responsible for the different outcomes obtained from this study. With this sensitivity and specificity, in-house PCR method which we used is a candidate for routine diagnosis.

摘要

两种最常见的人类多瘤病毒是BK病毒(BKV)和JC病毒(JCV)。与多瘤病毒相关的疾病通常发生在严重细胞免疫抑制的情况下。BKV和JCV可引起多种疾病,特别是在免疫抑制宿主中重新激活时。本研究的目的是比较和评估针对病毒基因组小T和大T基因区域的实时聚合酶链反应(PCR)方法的结果,同时考虑BKV和JCV病毒基因组中出现的多态性。 从免疫抑制患者中采集了82例患者的尿液标本,并送至梅拉姆医学院分子微生物学实验室。使用商业试剂盒(LightMix,罗氏)通过实时PCR方法研究小t基因。使用优化的内部实时PCR方法研究大T基因。序列分析被视为标准方法。 通过针对小t基因区域的实时PCR方法,在9个样本中检测到BKV阳性,在61个样本中检测到JCV阳性;通过针对大T基因区域的实时PCR方法,在21个样本中确定BKV阳性,在67个样本中确定JCV阳性。统计学上,两种方法之间BKV检测存在显著差异,而JCV检测无显著差异。 目标基因区域的不同多态性导致了本研究获得的不同结果。基于这种敏感性和特异性,我们使用的内部PCR方法是常规诊断的候选方法。

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