基于Taq Man探针的四重实时荧光定量PCR检测甲型/乙型/丙型副伤寒沙门菌和伤寒沙门菌
[Taq Man probe-based quadruple real-time PCR for detection of Salmonella paratyphi A/B/C and Salmonella typhi].
作者信息
Xu Longyan, Yuan Muyun, Sun Wei, Ke Bixia, Hong Ye
机构信息
Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou 510623, China.
出版信息
Wei Sheng Yan Jiu. 2017 Mar;46(2):298-302.
OBJECTIVE
Establishment and application of Taq Man probe-based quadruple real-time PCR for detection of Salmonella paratyphi A/B/C and Salmonella typhi. Primers specific to Salmonella paratyphi A( SPAP), Salmonella paratyphi B( SPBP), Salmonella paratyphi C( SPCP), and Salmonella typhi( STP) were designed.
METHODS
A method of Taq Man probe-based quadruple real-time PCR was established according to the distinction of the 5'end of the probe mark of TET, ROX, FAM and HEX. 5 strains of SPA, 4 strains of SPB, 7 strains of SPC and 11 strains of ST were identified by amplification from SPAP, SPBP, SPCP and STP.
RESULTS
While other serotypes of salmonella and17 strains of non-salmonella got negative results of amplification. Amplification rate of SPAP, SPBP, SPCP, and STP were 84. 5%, 101. 8%, 92. 4% and 90. 9%, respectively. The linear correlation coefficient( R~2) were 0. 996, 0. 975, 0. 996 and 0. 984, respectively.
CONCLUSION
The PCR system is specific and sensitive for the identification of SPA, SPB, SPC and ST.
目的
建立并应用基于Taq Man探针的四重实时荧光定量PCR检测甲型/乙型/丙型副伤寒沙门菌及伤寒沙门菌。设计针对甲型副伤寒沙门菌（SPAP）、乙型副伤寒沙门菌（SPBP）、丙型副伤寒沙门菌（SPCP）和伤寒沙门菌（STP）的特异性引物。
方法
根据探针5′端标记TET、ROX、FAM和HEX的不同，建立基于Taq Man探针的四重实时荧光定量PCR方法。通过SPAP、SPBP、SPCP和STP引物对5株甲型副伤寒沙门菌、4株乙型副伤寒沙门菌、7株丙型副伤寒沙门菌和11株伤寒沙门菌进行扩增鉴定。
结果
其他血清型沙门菌及17株非沙门菌扩增结果均为阴性。SPAP、SPBP、SPCP和STP的扩增率分别为84.5%、101.8%、92.4%和90.9%。线性相关系数（R²）分别为0.996、0.975、0.996和0.984。
结论
该PCR体系对甲型副伤寒沙门菌、乙型副伤寒沙门菌、丙型副伤寒沙门菌和伤寒沙门菌的鉴定具有特异性和敏感性。
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