Zuo Le, Jiang Yixiang, Wan Min, Jiang Min, Shi Xiaolu, Li Yinghui, Qiu Yaqun, Lin Yiman, Hu Qinghua
Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, China.
Wei Sheng Yan Jiu. 2020 Sep;49(5):823-858. doi: 10.19813/j.cnki.weishengyanjiu.2020.05.022.
Multiplex real-time PCR for the identification of 15 Salmonella serovars was developed.
Through the Salmonella genome comparison, 12 membrane proteins STM4497 gene can be used to identify 15 Salmonella serovars, and these 12 genes were respectively listed as A-L genes. Then primers were designed according to A-L gene conserved sequences, and then multiplex real-time PCR was established assessed with the evaluation of the limit detection, sensitivity, specificity, and repeatability. The 206 Salmonella strains were identified using multiplex real-time PCR with the comparison of the serum slide agglutination assay.
The limit detection of multiplex PCR ranged from 1. 1×10(-3)-1. 2×10(-3) ng/μL. The target genes were 100% specificity, and the relative standard deviation was lower than 2. 97%. Compared with the serum slide agglutination assay, Kappa ranged 0. 92-1. 00.
The multiplex real-time PCR can be used to identify 15 Salmonella serovars, which is rapid, accurate and specific.
建立用于鉴定15种沙门氏菌血清型的多重实时荧光定量PCR方法。
通过沙门氏菌基因组比较，筛选出可用于鉴定15种沙门氏菌血清型的12个膜蛋白STM4497基因，分别命名为A-L基因。根据A-L基因保守序列设计引物，建立多重实时荧光定量PCR方法，并对其检测限、灵敏度、特异性和重复性进行评价。采用多重实时荧光定量PCR对206株沙门氏菌进行鉴定，并与血清玻片凝集试验结果进行比较。
多重实时荧光定量PCR检测限为1.1×10(-3)-1.2×10(-3) ng/μL。目的基因特异性为100%，相对标准偏差低于2.97%。与血清玻片凝集试验比较，Kappa值为0.92-1.00。
该多重实时荧光定量PCR方法可用于鉴定15种沙门氏菌血清型，具有快速、准确、特异的特点。
