Detection of Yersinia pestis DNA in Prairie Dog-Associated Fleas by Polymerase Chain Reaction Assay of Purified DNA
Detection of Yersinia pestis DNA in Prairie Dog-Associated Fleas by Polymerase Chain Reaction Assay of Purified DNA
复制标题
DOI：
10.7589/0090-3558-46.2.636
复制
发表时间：
2010-04-01
影响因子：
1.3
通讯作者：
Miller, Michael W.
中科院分区：
文献类型：
--
作者：
Griffin, Karen A.;Martin, Daniel J.;Miller, Michael W.
我们评估，完善，并应用完善的聚合酶链反应（PCR）技术检测鼠疫耶尔森菌。从土拨鼠（土拨鼠）洞穴中采集的跳蚤（主要是土拨鼠）的DNA。根据鼠疫杆菌A1122毒株DNA的PCR结果，采用DNA纯化和纤溶酶原激活物基因引物对野外采集的蚤进行筛选。我们在两个黑尾草原土拨鼠（Cynomys ludovicianus）群落的跳蚤池中检测到鼠疫耶尔森氏菌DNA，这些群落有证据表明最近发生了鼠疫兽疫，而在四个白尾草原土拨鼠（Cynomys leucurus.）群落复合体（Wolf Creek）中检测到鼠疫耶尔森氏菌DNA缺乏兽疫证据。在狼溪，跳蚤池中的相对跳蚤丰度和鼠疫杆菌DNA的发生似乎随时间而变化。在未观察到动物鼠疫的情况下，DNA纯化和引物序列似乎都影响了PC R法检测鼠疫耶尔森菌DNA的可能性。在Wolf Creek草原木屐洞穴中收集的跳蚤中存在鼠疫杆菌质粒DNA，可能表明受感染的跳蚤在动物流行病之间以某种方式维持在该系统中，这与鼠疫在其他地方草原木屐群落复合体中保持地方性的假设一致。
We evaluated, refined, and applied well-established polymerase chain reaction (PCR) techniques for detecting Yersinia pestis. DNA in fleas (mainly Oropsylla spp.) collected from prairie dog (Cynomys spp.) burrows. Based on results from PCR of avirulent Y. pestis strain A1122 DNA, we used DNA purification and primers for the plasminogen activator gene to screen field-collected fleas. We detected Y. pestis DNA in flea pools from two black-tailed prairie dog (Cynomys ludovicianus) colonies with evidence of recent plague epizootics, and from one of four white-tailed prairie clog (Cynomys leucurus.) colony complexes (Wolf Creek) where evidence of epizootic plague was lacking. Relative flea abundance and occurrence of Y. pestis DNA among flea pools appeared to vary over time at Wolf Creek. Both DNA purification and primer sequences appeared to influence the likelihood of detecting Y. pestis DNA by PC R in fleas collected from prairie clog burrows in the absence of observed epizootic plague. Presence of Y. pestis plasmid DNA in fleas collected from prairie clog burrows at Wolf Creek may represent evidence that infected fleas were somehow being maintained in that system between epizootics, consistent with the hypothesized enzootic maintenance of plague in prairie clog colony complexes elsewhere.