Targeted PCR Rickettsia screen

A targeted PCR <i>Rickettsia </i>screen of 1,612 individuals from 169 species was undertaken to increase our understanding of <i>Rickettsia</i> 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 <i>Rickettsia </i>symbiosis (water beetles, biting midges, damselflies) reflects a wider higher incidence in species from this habitat. <br>mtDNA <i>COI </i>amplification was conducted as a control for DNA quality. To investigate symbiont infection status, rickettsial-specific primers based on<i> gltA</i> and <i>16S rRNA </i>genes were used for conventional PCR screening, with Sanger sequences obtained from at least one specimen per <i>Rickettsia </i>positive species to identify any misamplification false positives. <br>

为加深对立克次体属（*Rickettsia*）生态学的认知，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）筛查。本次筛查涵盖水生与陆生类群，旨在验证此前研究将特定水生类群（水生甲虫、蠓类、豆娘）认定为立克次体共生热点的结论，是否反映出该生境物种中该共生体的感染率普遍更高。
本研究以线粒体DNA细胞色素c氧化酶亚基I（mtDNA COI）扩增作为DNA质量质控手段。
为探究共生体感染状态，本研究采用基于*gltA*与*16S rRNA*基因的立克次体特异性引物开展常规PCR筛查，并对每一份立克次体检测阳性物种的至少1份标本进行桑格（Sanger）测序，以排查非特异性扩增导致的假阳性结果。