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《Zhejiang Journal of Preventive Medicine》 2013-01
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Developed Method of a Multiplex PCR Assay for the Identification of Salmonella Enterica Serovar Typhi

WEI Yi-cheng,ZHAN Li,YE Ju-lian.Center for Disease Control and Prevention of Pan' An,Pan' An,Zhejiang,322300,China  
Objective To develop a multiplex PCR assay for the detection of Salmonella enterica Serovar Typhi.Methods According to the gene sequences of somatic antigen for salmonella serogroups A/D1,fliC-Hd and Salmonella Vi capsular antigen(Vi),four pairs primers were designed.The multiplex PCR was developed and optimized.To valid the assay,genomic DNA from 15 salmonella strains representing 15 serotypes and 18 non-salmonella strains was subjected to the PCR.The method was applied to the detection of 50 samples isolated in Zhejiang Province.Results A multiple PCR was established to detect Salmonella enterica Serovar Typhi.The optimized reaction mixture(25 μl total volume) contained 100 μM dNTP mix,0.2 μM of each primer,2.5 U Taq DNA polymerase and 5 μl template.The cycling parameters of the multiplex PCR consisted of denaturation at 94 ℃ for 1 min,followed by 35 cycles at 94 ℃ for 1 min,56 ℃ for 1 min,72 ℃ for 1 min.This assay can differentiate Salmonella serogroup A from Salmonella serogroup D.and it also can identify the salmonella with fliC-Hd and the salmonella with Vi capsular antigen.The coincidence rate of the actual sample detection is up to 100.00%.Conclusion The multiplex PCR assay is highly selective and sensitive in detecting Salmonella Typhi and can be used as a supplementary method to traditional serum agglutination test.
【Fund】: 浙江省医药卫生平台骨干人才计划(2011RCB014)
【CateGory Index】: R378
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