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改良环介导等温扩增试剂盒在检测诺如病毒 I 型和 II 型中的应用。

Application of a modified loop-mediated isothermal amplification kit for detecting Norovirus genogroups I and II.

机构信息

Department of Infectious Diseases, Osaka Prefectural Institute of Public Health, Osaka, Japan.

出版信息

J Med Virol. 2009 Dec;81(12):2072-8. doi: 10.1002/jmv.21626.

Abstract

Norovirus is a major etiologic agent in worldwide outbreaks of gastroenteritis associated with food as well as person-to-person transmission. The ubiquitous nature of Norovirus necessitates simple and rapid detection methods with high accuracy and sensitivity. To this end, several investigators have evaluated the usefulness of commercial reverse-transcription loop-mediated isothermal amplification (RT-LAMP) kits for detecting Norovirus genogroups I (GI) and II (GII). In previous studies, the conventional Loopamp kit for Norovirus GII showed a relatively high detection rate, while that for Norovirus GI showed a relatively low detection rate. In the present study, clinical Norovirus specimens were used to compare the detection rate of a modified Loopamp kit for Norovirus GI with the rates of the conventional Loopamp kit for Norovirus GI and an "in-house" RT-LAMP GI primer set, methods which had a high detection rate. Results from the present study showed that the modified Loopamp kit for Norovirus GI had a higher detection rate for two viral genotypes (GI.3, GI.11). On comparison with an "in-house" GII primer set using genotype GII.4 viruses circulating recently, the detection rate by the Loopamp kit for Norovirus GII was found to be higher, with a 98% detection rate. These results indicate the applicability of the modified LAMP kit for GI and the conventional LAMP kit for GII for detection of Noroviruses in clinical samples.

摘要

诺如病毒是导致全球食源性和人际传播胃肠炎暴发的主要病原体。由于诺如病毒无处不在,因此需要使用简单、快速、准确且灵敏的检测方法。为此,已有多位研究人员评估了商用逆转录环介导等温扩增(RT-LAMP)试剂盒在检测诺如病毒 I 型(GI)和 II 型(GII)中的应用价值。在之前的研究中,常规的 Norovirus GII Loopamp 试剂盒显示出相对较高的检测率,而 Norovirus GI Loopamp 试剂盒的检测率则相对较低。在本研究中,我们使用临床 Norovirus 标本比较了改良的 Norovirus GI Loopamp 试剂盒与常规 Norovirus GI Loopamp 试剂盒和高检测率的“内部”RT-LAMP GI 引物组的检测率。本研究结果表明,改良的 Norovirus GI Loopamp 试剂盒对两种病毒基因型(GI.3、GI.11)的检测率更高。与最近流行的基因型 GII.4 病毒的“内部”GII 引物组相比,Norovirus GII Loopamp 试剂盒的检测率更高,达到 98%。这些结果表明,改良的 LAMP 试剂盒可用于 GI 的检测,而常规的 LAMP 试剂盒可用于 GII 的检测,可用于临床样本中诺如病毒的检测。

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