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一种基于高分辨率熔解(HRM)技术的检测方法,用于单纯疱疹病毒1型(HSV-1)和单纯疱疹病毒2型(HSV-2)的经济高效临床检测和基因分型。

A high resolution melting (HRM) technology-based assay for cost-efficient clinical detection and genotyping of herpes simplex virus (HSV)-1 and HSV-2.

作者信息

Lieveld M, Carregosa A, Benoy I, Redzic N, Berth M, Vanden Broeck D

机构信息

International Centre for Reproductive Health, Department of Urogynaecology, Ghent University Hospital, Ghent, Belgium.

Algemeen Medisch Laboratorium, Sonic Healthcare, Antwerp, Belgium.

出版信息

J Virol Methods. 2017 Oct;248:181-186. doi: 10.1016/j.jviromet.2017.07.005. Epub 2017 Jul 19.

Abstract

Genital herpes can be caused by two very similar viruses, herpes simplex virus (HSV)-1 or HSV-2. These two HSV types cannot be distinguished clinically, but genotyping is recommended in the first-episodes of genital herpes to guide counselling and management. Quantitative polymerase chain reaction (qPCR) is the preferred diagnostic method for HSV typing. However, commercial qPCR methods use expensive fluorescent labeled probes for detection. Furthermore, most low-cost methods are not able to differentiate between HSV-1 and -2. The aim of this study was to develop a high resolution melting (HRM) technology-based assay for sensitive HSV-1 and HSV-2 detection and genotyping. Using a panel of 46 clinical specimens, the performance of the HRM assay was compared to two commercial HSV tests: the HRM assay detected HSV in all 23 positive samples, with no false positive results (100% concordance with HSV I/II Real-TM assay). Additionally, the HRM assay correctly genotyped both HSV types in a subset of these clinical samples, as determined by the Realstar HSV PCR Kit. The HSV HRM assay provides a cost-effective alternative method to conventional more expensive assays and can be used in routine clinical specimens, in cases where it is particularly necessary to detect and distinguish HSV-1 from -2.

摘要

生殖器疱疹可由两种非常相似的病毒引起,即单纯疱疹病毒(HSV)-1或HSV-2。这两种HSV类型在临床上无法区分,但在生殖器疱疹初发时建议进行基因分型,以指导咨询和管理。定量聚合酶链反应(qPCR)是HSV分型的首选诊断方法。然而,商业qPCR方法使用昂贵的荧光标记探针进行检测。此外,大多数低成本方法无法区分HSV-1和-2。本研究的目的是开发一种基于高分辨率熔解(HRM)技术的检测方法,用于敏感地检测HSV-1和HSV-2并进行基因分型。使用一组46份临床标本,将HRM检测方法的性能与两种商业HSV检测方法进行了比较:HRM检测方法在所有23份阳性样本中均检测到HSV,无假阳性结果(与HSV I/II Real-TM检测方法100%一致)。此外,根据Realstar HSV PCR试剂盒的检测结果,HRM检测方法在这些临床样本的一个子集中正确地对两种HSV类型进行了基因分型。HSV HRM检测方法为传统的更昂贵的检测方法提供了一种经济有效的替代方法,可用于常规临床标本,特别是在需要检测和区分HSV-1和-2的情况下。

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