基于TaqMan探针四重荧光PCR检测甲型、乙型、丙型副伤寒和伤寒沙门菌

发布时间：2018-03-16 15:21

本文选题：四重荧光PCR　切入点：甲型副伤寒沙门菌　出处：《卫生研究》2017年02期 　论文类型：期刊论文

【摘要】：目的建立基于Taq Man探针四重荧光PCR检测甲型、乙型、丙型副伤寒和伤寒沙门菌的方法。方法根据甲型副伤寒沙门菌(Salmonella paratyphi A,SPA)、乙型副伤寒沙门菌(Salmonella paratyphi B,SPB)、丙型副伤寒沙门菌(Salmonella paratyphi C,SPC)和伤寒沙门菌(Salmonella typhi,ST)的特异序列分别设计引物SPAP、SPBP、SPCP和STP,在探针的5'端分别标记TET、ROX、FAM、HEX,建立基于Taq Man探针四重荧光PCR检测方法。结果 SPAP、SPBP、SPCP和STP分别扩增出5株SPA、4株SPB、7株SPC和11株ST,而其他血清型沙门菌和17株非沙门菌扩增结果阴性。SPAP、SPBP、SPCP和STP扩增效率分别为84.5%、101.8%、92.4%和90.9%,线性相关系数(R~2)分别为0.996、0.975、0.996和0.984。结论建立的方法特异性高、敏感性强,可用于SPA、SPB、SPC和ST的特异性检测。
[Abstract]:Objective to establish a quadruplex fluorescence PCR based on Taq Man probe for the detection of A and B type. Methods the specific sequences of Salmonella paratyphi, Salmonella paratyphi, Salmonella paratyphi, Salmonella paratyphi paratyphi and Salmonella typhimurium Typhoid Salmonella typhimurium Typhoid were designed and introduced by using the specific sequences of Salmonella paratyphi, Salmonella paratyphi, Salmonella paratyphi, Salmonella paratyphi, and Salmonella typhimurium, respectively, of Salmonella paratyphoid A and Salmonella typhimurium. SPAPPP-SPBPP-SPCP and STP were labeled at the 5'end of the probe, respectively, and a method was established for the detection of SPPAB7 SPC and 11 STs based on Taq Man probe. The results showed that 5 strains of spa, 4 strains of spa, 7 strains of SPC and 11 strains of STP were amplified respectively, while other serotype Salmonella and 17 strains of non-salmonella were amplified. The amplification efficiency of SPCP and STP were 84.5% and 90.9%, respectively, and the linear correlation coefficient was 0.996 and 0.984.Conclusion the specificity of the method was high. It is sensitive and can be used for the specific detection of SPC and St in spa.
【作者单位】：广东检验检疫技术中心;广州机场检验检疫局;广东省疾病预防控制中心;
【基金】：广东省科技项目计划(No.2013B040402004)
【分类号】：R440;R516.3

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