Department of Molecular Microbiology, Hospital Universitari Sant Joan de Deu, Esplugues, Barcelona, Spain.
Eur J Clin Microbiol Infect Dis. 2012 May;31(5):715-9. doi: 10.1007/s10096-011-1364-1. Epub 2011 Jul 31.
Molecular detection of enterovirus (EV) RNA based on PCR methods is a quicker and more sensitive approach than culture methods. At present, different PCR-based methods for EV RNA detection are available, but comparisons of results obtained according to the different approaches are limited. We evaluated an in-house real-time RT-PCR assay with a commercialized TaqMan real-time RT-PCR kit for detection of EV. Consecutive clinical specimens from paediatric patients less than 6 years old with clinical suspicion of EV infection were analyzed between July and November 2010. After RNA extraction, samples were amplified both by the real-time RT-PCR commercial assay and the in-house assay. A total of 19 of 132 patients (14.4%) involving 20 samples (14 plasma samples and 6 CSF) were positive in at least one of the two assays. The sensitivity of the in-house assay when the MutaPLATE® assay was used as a reference was 90% (IC 95%; 74.35-100) and the specificity was 100% (IC 95%; 99.63-100). Cts results of two methods were statistically correlated (r = 0.774; P = 0.01). In conclusion, these two real-time RT-PCR assays are rapid and easy methods for detection of EV.
基于 PCR 方法的肠道病毒 (EV) RNA 的分子检测比培养方法更快、更敏感。目前,有多种基于 PCR 的 EV RNA 检测方法,但根据不同方法获得的结果比较有限。我们评估了一种用于 EV 检测的内部实时 RT-PCR 检测法和商业化的 TaqMan 实时 RT-PCR 试剂盒。2010 年 7 月至 11 月,对临床怀疑感染 EV 的 132 例年龄小于 6 岁的儿科患者连续采集临床标本进行分析。提取 RNA 后,通过实时 RT-PCR 商业检测法和内部检测法对样本进行扩增。在这两种检测方法中,至少有一种检测方法呈阳性的患者共有 19 例(14.4%),涉及 20 份样本(14 份血浆样本和 6 份脑脊液样本)。当以 MutaPLATE®检测法作为参考时,内部检测法的灵敏度为 90%(95%CI;74.35-100),特异性为 100%(95%CI;99.63-100)。两种方法的 Cts 结果具有统计学相关性(r = 0.774;P = 0.01)。总之,这两种实时 RT-PCR 检测法是检测 EV 的快速简便方法。