ISSN 1003-8280 CN 10-1522/R 中国疾病预防控制中心 主办
Chinese Journal of Vector Biology and Control >
Development of PCR-RFLP for rapid differentiation of Yersinia pestis strains between Microtus and non-Microtus biotypes in China
Received date: 2014-07-25
Online published: 2014-12-20
Supported by
Supported by the Special Fund for Health Sector, People's Republic of China (No. 201202021)
Objective To develop a simple genotyping method for rapid differentiation of Yersinia pestis strains between Microtus and non-Microtus biotypes. Methods Specific primers were designed and applied to amplify the aspartase gene (aspA) through polymerase chain reaction (PCR) among 93 Y. pestis strains of diverse origins and biotypes. The PCR products were genotyped by restriction fragment length polymorphism (RFLP) analysis using the restriction enzyme Hpy CH4Ⅳ. Genetic polymorphisms of aspA were determined by direct DNA sequencing. Results Among the 93 test strains, the amplified products of aspA digested with Hpy CH4Ⅳ showed two genotypes: Microtus biotype strains (38) all had two RFLP bands in sizes of 101 and 126 bp, respectively; and non-Microtus biotype strains (58) consistently had one single RFLP band in size of 227 bp. Direct sequencing data confirmed that mutation at the recognition site for restriction enzyme led to differences in the RFLP banding patterns. Conclusion A new genotyping method (aspA-PCR-RFLP) was established for rapid differentiation of Y. pestis strains between Microtus biotype with low virulence and non-Microtus biotype with high virulence according to different RFLP banding patterns. The proposed method can be used for epidemiologic surveillance in plague foci.
ZHENG Xiao, XIA Lian-xu, HAI Rong, ZHANG Zhi-kai, CAI Hong, YOU Xin, PING Jing, LI Wei . Development of PCR-RFLP for rapid differentiation of Yersinia pestis strains between Microtus and non-Microtus biotypes in China[J]. Chinese Journal of Vector Biology and Control, 2014 , 25(6) : 489 -491 . DOI: 10.11853/j.issn.1003.4692.2014.06.001
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