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在逆转录聚合酶链反应分析中使用NSP3基因区分罗特威疫苗株与野生型株。

Differentiation of RotaTeq vaccine strains from wild-type strains using NSP3 gene in reverse transcription polymerase chain reaction assay.

作者信息

Jeong Sunyoung, Than Van Thai, Lim Inseok, Kim Wonyong

机构信息

Department of Microbiology, Chung-Ang University College of Medicine, Seoul, South Korea.

Department of Pediatrics, Chung-Ang University College of Medicine, Seoul, South Korea.

出版信息

J Virol Methods. 2016 Nov;237:72-78. doi: 10.1016/j.jviromet.2016.08.022. Epub 2016 Aug 30.

Abstract

RotaTeq is a live attenuated human-bovine reassortant vaccine against rotaviruses that is used worldwide. However, shedding of the virus used in RotaTeq has been detected in the feces of children following vaccination by the oral route, possibly affecting community immunity. Therefore, a simple and efficient method to discriminate between virulent and RotaTeq vaccine strains is required. In this study, a novel one-step multiplex reverse-transcription polymerase chain reaction (RT-PCR) assay targeting the NSP3 gene was developed to detect RotaTeq vaccine strains in fecal samples. RotaTeq vaccine viruses were successfully distinguished from known wild-type rotavirus genotypes. In addition, the developed assay was able to detect rotaviruses in clinical stool samples obtained from South Korea during the 2011-2013 rotavirus seasons. Of the 1106 stool specimens from children with acute gastroenteritis that were screened, 286 rotaviruses were genotyped. RotaTeq vaccine strains were identified in 39 samples (13.6%). The novel RT-PCR assay that was developed could be used to detect and discriminate between RotaTeq vaccine strains that are shed in fecal matter, and to estimate the quantification of virus that has been shed after vaccination.

摘要

Rotateq是一种用于全球范围的针对轮状病毒的人-牛重组减毒活疫苗。然而,口服接种Rotateq后,在儿童粪便中检测到了该疫苗所用病毒的排出,这可能会影响群体免疫。因此,需要一种简单有效的方法来区分强毒株和Rotateq疫苗株。在本研究中,开发了一种针对NSP3基因的新型一步多重逆转录聚合酶链反应(RT-PCR)检测方法,以检测粪便样本中的Rotateq疫苗株。Rotateq疫苗病毒成功地与已知的野生型轮状病毒基因型区分开来。此外,所开发的检测方法能够检测2011 - 2013年轮状病毒流行季节期间从韩国获得的临床粪便样本中的轮状病毒。在对1106份急性胃肠炎儿童粪便标本进行筛查后,对286份轮状病毒进行了基因分型。在39份样本(13.6%)中鉴定出Rotateq疫苗株。所开发的新型RT-PCR检测方法可用于检测和区分粪便中排出的Rotateq疫苗株,并估计接种疫苗后排出病毒的数量。

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