通过巢式多重聚合酶链反应对疑似伤寒血样中伤寒沙门氏菌进行分子诊断及其毒力分析

Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR.

作者信息

Prabagaran Solai Ramatchandirane, Kalaiselvi Vellingiri, Chandramouleeswaran Naganathan, Deepthi Krishnan Nair Geetha, Brahmadathan Kootallur Narayanan, Mani Mariappa

机构信息

Department of Biotechnology, Bharathiar University, Coimbatore, Tamilnadu 641 046, India.

出版信息

J Microbiol Methods. 2017 Aug;139:150-154. doi: 10.1016/j.mimet.2017.05.013. Epub 2017 May 23.

DOI:10.1016/j.mimet.2017.05.013

PMID:28545919

Abstract

A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typhi. Among them, those positive for fliC alone were subjected to identification of genes in Via B operon of Salmonella Pathogenesity Island (SPI-7) where four primer pairs were used to detect tviA and tviB genes. Among 250 blood samples tested, 115 were positive by fliC PCR; 22 of these were negative for tviA and tviB. Hence, the method described here can be used to diagnose the incidence of Vi-negative serovar typhi especially in endemic regions where the Vi vaccine is administered.

摘要

开发了一种基于巢式多重聚合酶链反应（PCR）的诊断方法，用于检测疑似伤寒热患者血液样本中的强毒力伤寒沙门氏菌。在维达试验之后，使用两对引物检测伤寒沙门氏菌的鞭毛蛋白基因（fliC）。其中，仅fliC呈阳性的样本进行沙门氏菌致病岛（SPI-7）的Via B操纵子中的基因鉴定，使用四对引物检测tviA和tviB基因。在检测的250份血液样本中，115份通过fliC PCR呈阳性；其中22份tviA和tviB呈阴性。因此，这里描述的方法可用于诊断Vi阴性伤寒血清型的发病率，特别是在接种Vi疫苗的流行地区。