应用实时荧光定量PCR检测鼠类生境样本鼠疫耶尔森氏菌的研究

Detection of Yersinia pestis from mice habitat samples by real-time PCR

目的评价应用实时荧光定量PCR(RT-PCR)方法检测鼠类生境样本中鼠疫耶尔森氏菌的可行性和有效性。方法优选、评价鼠类生境样本DNA提取方法,提取疫区鼠类洞穴土壤、粪便等生境样本中DNA,应用RT-PCR试剂盒检测鼠疫耶尔森氏菌特异性pla基因。结果柱式土壤DNAout法提取DNA效果最好,回收率是24.54%。联合应用柱式土壤DNAout法和RT-PCR法检测鼠类生境样本,鼠疫耶尔森氏菌pla基因的阳性率为3.44%(10/291)。结论应用柱式土壤DNAout法提取鼠类生境样本DNA,RT-PCR法可以检测到鼠类生境中鼠疫耶尔森氏菌pla基因。

Objective To evaluate the feasibility and validity of detecting Yersinia pestis from mice habitat samples by RT-PCR.Methods The different methods of collecting DNA from mice habitat samples were optimized and evaluated.DNA from plague epidemic area samples such as soil,feces in the mice caves was collected by using the best method,and pla gene of Yersinia pestis was detected with RT-PCR Kit.Results The column soil DNAout method was the most effective to collect DNA from mice habitat samples,the rate of recovery was 24.54%.Combined this method with RT-PCR to detect the mice habitat samples from plague epidemic area,the positive rate of Yersinia.pestis pla gene was 3.44%（10/291）.Conclusions Using column soil DNAout method to collect DNA from mice habitat samples,the pla gene of Yersinia pestis among the mice habitat samples can be detected by RT-PCR.