Development of a Rickettsia bellii-Specific TaqMan Assay Targeting the Citrate Synthase Gene
Development of a Rickettsia bellii-Specific TaqMan Assay Targeting the Citrate Synthase Gene
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DOI：
10.1093/jme/tjw111
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发表时间：
2016-11-01
影响因子：
2.1
通讯作者：
Karpathy, Sandor E.
中科院分区：
文献类型：
--
作者：
Hecht, Joy A.;Allerdice, Michelle E. J.;Karpathy, Sandor E.
贝氏立克次体是一种致病性未知的立克次体，感染美洲各地的硬蜱和硬蜱。许多用于检测斑点热群（SFG）立克次体的分子检测法不能检测到立克次体。因此，当使用特异性筛选SFG立克次体的检测方法时，这种细菌的感染可能隐藏在蜱虫种群中。我们描述了一个R。bellii特异性、定量、实时PCR TaqMan测定，其靶向柠檬酸合酶（gltA）基因的片段。针对一组DNA样本验证了该检测方法的特异性，其中包括26种立克次体、东方体、埃立克体、无形体和巴尔通体，5种蜱和人DNA样本，以及20种立克次体分离株的DNA。贝利，包括11个来自北美和9个来自南美。阿尔河构建bellii对照质粒，并使用质粒的系列稀释液确定测定的检测限为每4ml模板DNA一个拷贝。该方法可用于更好地确定R.贝利在西半球蜱传立克次体病流行病学中的作用。
Rickettsia bellii is a rickettsial species of unknown pathogenicity that infects argasid and ixodid ticks throughout the Americas. Many molecular assays used to detect spotted fever group (SFG) Rickettsia species do not detect R. bellii, so that infection with this bacterium may be concealed in tick populations when assays are used that screen specifically for SFG rickettsiae. We describe the development and validation of a R. bellii-specific, quantitative, real-time PCR TaqMan assay that targets a segment of the citrate synthase (gltA) gene. The specificity of this assay was validated against a panel of DNA samples that included 26 species of Rickettsia, Orientia, Ehrlichia, Anaplasma, and Bartonella, five samples of tick and human DNA, and DNA from 20 isolates of R. bellii, including 11 from North America and nine from South America. A R. bellii control plasmid was constructed, and serial dilutions of the plasmid were used to determine the limit of detection of the assay to be one copy per 4 ml of template DNA. This assay can be used to better determine the role of R. bellii in the epidemiology of tickborne rickettsioses in the Western Hemisphere.