Using real-time fluorescence quantitative PCR method to rapidly differentiate Microtus and non-Microtus Yersinia pestis
Objective To establish a real-time fluorescence quantitative PCR method for rapid differentiating the microtus and non-microtus Yersinia pestis.Methods The aspartase gene(aspA)of Yersinia pestis is pathogen-related gene,and the allelic gene mutation(G/T)at 1087 base pair site of the asp A gene can effectively distinguish the microtus and non-microtus Yersinia pestis.Based on the allelic gene mutation site,specific differentiating probes and primers were designed to establish a fluorescence quantitative PCR method to identigy microtus and non-microtus Yersinia pestis.The discriminatory effectiveness of this method was evaluated by testing representative strains from plague foci in China.Results The established specific differential probe fluorescence quantitative PCR method can accurately distinguish the microtus and non-microtus Yersinia pestis,with no amplification observed in non-Yersinia pestis strains.Conclusion This method can be used to differentiate the microtus and non-microtus Yersinia pestis strains,making it suitable to identify surveillance strains quickly from plague foci in China.
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