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

    潘光合1，梁亮2，万道正2，梁中平2，杨会敏2，于建国1，田珍灶1，蓝翊文2

    1.广西国际旅行卫生保健中心，广西 南宁 530021；2.凭祥出入境检验检疫局

    摘要：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。

    关键词：鼠疫耶尔森氏菌；鼠；洞穴；生境；实时荧光定量PCR

    中图分类号：R184.35　文献标识码： B

    Detection of Yersinia pestis from micehabitat samples by real-time PCR

    PAN Guang-he*, LIANG Liang, WAN Dao-zheng,LIANG Zhong-ping, YANG Hui-min,

    YU Jian-guo, TIAN Zhen-zao, LAN Yi-wen

    *Guangxi International Travel Hearth CareCenter, Nanning, Guangxi 530021,China

    Abstract:   Objective  To evaluate the feasibility and validity of detecting Yersiniapestis from mice habitat samples by RT-PCR. Methods   The different methods of collecting DNA frommice habitat samples were optimized and evaluated. DNA from plagueepidemic area samples such as soil,feces in the mice caves wascollected by using the best method, and pla gene of Yersinia pestiswas detected with RT-PCR Kit.  Results   The columnsoil DNAout method was the most effective to collect DNA from micehabitat samples, the rate of recovery was 24.54%. Combined thismethod with RT-PCR to detect the mice habitat samples from plagueepidemic area, the positive rate of Yersinia. pestis pla gene was3.44% (10/291).  Conclusions   Using column soilDNAout method to collect DNA from mice habitat samples ,the plagene of Yersinia pestis among the mice habitat samples can bedetected by RT-PCR.

    Key words:   Yersinia pestis；Mice；Burrrow ；Habitat；RT-PCR