Establishment and application of a genotyping technique for detection of different DNA regions in Yersinia pestis using multiple polymerase chain reaction

Abstract

Objective By the method of multiple polymerase chain reaction (PCR), we intend to amplify different regions (DFR) of Yersinia pestis DNA, and to establish a multiple DFR genotyping technique for detection of Yersinia pestis. Methods According to the product size of 23 DFRs and pMT plasmid, 24 primers were optimized and combined, then multiple primers in one PCR reaction system were added, and positive template DNA was amplified. Meanwhile, 200 wild strain DNAs were amplified by multiple PCR and normal PCR, to verify the coincidence rate of the two methods. Results Totally 24 target segments were amplified through the positive DNA template. Through different permutation and combination, 24 primers were optimized and combined into 9 groups. Totally 200 wild strain DNAs were used for verification, the coincidence rate of multiple PCR and normal PCR was 100%. Conclusions Multiple PCR is applicable and feasible for DFR genotyping of Yersinia pestis. It is an efficient, economic and high accuracy experimental method for large quantities of Yersinia pestis DFR genotyping. Key words: Multiple polymerase chain reaction; Yersinia pestis; Different region