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首页> 外文期刊>The Journal of molecular diagnostics: JMD >Clinical Validation of a New Triplex Real-Time Polymerase Chain Reaction Assay for the Detection and Discrimination of Herpes simplex Virus Types 1 and 2.
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Clinical Validation of a New Triplex Real-Time Polymerase Chain Reaction Assay for the Detection and Discrimination of Herpes simplex Virus Types 1 and 2.

机译：一种用于检测和区分1型和2型单纯疱疹病毒的新型三重实时聚合酶链反应测定的临床验证。

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摘要

A new triplex real-time polymerase chain reaction (PCR) assay for Herpes simplex virus (HSV) (artus HSV-1/2 TM PCR kit, QIAGEN) was evaluated. This assay simultaneously uses three differently labeled probes targeted to HSV-1 (FAM), HSV-2 (NED), and to the manufacturer's Internal Control (VIC). HSV-1/2 typing capability and quantitation accuracy were determined using HSV stocks and quality control panels. Performance in routine clinical testing was compared with a nested HSV-1/2 PCR assay. Dilution series and quality control panel testing revealed an approximately 10-fold higher HSV-2 sensitivity in real-time PCR compared with an in-house nested PCR assay. The sensitivity for HSV-1 was comparable in both assays. All HSV-positive proficiency panel samples (n = 21) and virus stocks were typed correctly as HSV-1 or HSV-2 using real-time PCR. Quantitation correlated well with reference values (HSV-1, r = 0.98; HSV-2, r = 0.88), and 95% detection limits were determined as 9.4 HSV-2 copies/reaction and 18 HSV-1 copies/reaction. Based on C(t) values, the mean intra-assay coefficient of variation was 1%, whereas the interassay coefficient of variations were 2.7% and 2.5% for HSV-1 and -2, respectively. Testing of 309 clinical samples resulted in 100% specificity and 97% sensitivity. In conclusion, the artus HSV-1/2 TM PCR kit represents an excellent tool for the detection and differentiation of HSV-1 and -2 in clinical samples.
机译：评估了单纯疱疹病毒（HSV）的一种新的三重实时聚合酶链反应（PCR）检测方法（arts HSV-1 / 2 TM PCR kit，QIAGEN）。此测定法同时使用针对HSV-1（FAM），HSV-2（NED）和制造商内部控制（VIC）的三种不同标记的探针。 HSV-1 / 2分型能力和定量准确性使用HSV库存和质量控制面板确定。将常规临床测试中的性能与嵌套式HSV-1 / 2 PCR分析进行了比较。稀释系列和质量控制小组测试显示，与内部嵌套式PCR检测相比，实时PCR的HSV-2敏感性高约10倍。在两种试验中，HSV-1的敏感性均相当。使用实时PCR将所有HSV阳性熟练程度样本样本（n = 21）和病毒库正确键入为HSV-1或HSV-2。定量与参考值（HSV-1，r = 0.98; HSV-2，r = 0.88）很好地相关，测定95％的检出限为9.4 HSV-2拷贝/反应和18 HSV-1拷贝/反应。基于C（t）值，平均批内变异系数为1％，而HSV-1和-2批间变异系数分别为2.7％和2.5％。对309个临床样品进行测试得出的特异性为100％，灵敏度为97％。总之，Artus HSV-1 / 2 TM PCR试剂盒是检测和区分临床样品中HSV-1和-2的极佳工具。

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