Targeted PCR Rickettsia screen

A targeted PCR Rickettsia screen of 1,612 individuals from 169 species was undertaken to increase our understanding of Rickettsia ecology. Within this, we included a range of both aquatic and terrestrial taxa, to investigate if the previous work highlighting particular aquatic taxa as hotspots for Rickettsia symbiosis (water beetles, biting midges, damselflies) reflects a wider higher incidence in species from this habitat. mtDNA COI amplification was conducted as a control for DNA quality. To investigate symbiont infection status, rickettsial-specific primers based on gltA and 16S rRNA genes were used for conventional PCR screening, with Sanger sequences obtained from at least one specimen per Rickettsia positive species to identify any misamplification false positives.

为加深对立克次体（Rickettsia）生态学的理解，本研究针对169个物种的1612个个体开展了靶向聚合酶链式反应（PCR）立克次体筛查。本次筛查纳入了一系列水生与陆生分类群，旨在验证此前将特定水生分类群（水生甲虫、吸血蠓、豆娘）确定为立克次体共生热点的研究结论，是否反映了该生境物种普遍具有更高的立克次体感染率。本研究以线粒体DNA细胞色素C氧化酶亚基I（mtDNA COI）的扩增作为DNA质量质控对照。为探究共生菌感染状态，本研究采用基于gltA与16S核糖体RNA（16S rRNA）基因的立克次体特异性引物开展常规PCR筛查，并对每个立克次体阳性物种的至少1份标本进行桑格（Sanger）测序，以识别错扩导致的假阳性结果。