Massung R F, Slater K, Owens J H, Nicholson W L, Mather T N, Solberg V B, Olson J G
National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 1998 Apr;36(4):1090-5. doi: 10.1128/JCM.36.4.1090-1095.1998.
A sensitive and specific nested PCR assay was developed for the detection of granulocytic ehrlichiae. The assay amplifies the 16S rRNA gene and was used to examine acute-phase EDTA-blood and serum samples obtained from seven humans with clinical presentations compatible with human granulocytic ehrlichiosis. Five of the seven suspected cases were positive by the PCR assay using DNA extracted from whole blood as the template, compared with a serologic assay that identified only one positive sample. The PCR assay using DNA extracted from the corresponding serum samples as the template identified three positive samples. The sensitivity of the assay on human samples was examined, and the limit of detection was shown to be fewer than 2 copies of the 16S rRNA gene. The application of the assay to nonhuman samples demonstrated products amplified from template DNA extracted from Ixodes scapularis ticks collected in Rhode Island and from EDTA-blood specimens obtained from white-tailed deer in Maryland. All PCR products were sequenced and identified as specific to granulocytic ehrlichiae. A putative variant granulocytic ehrlichia 16S rRNA gene sequence was detected among products amplified from both the ticks and the deer blood specimens.
开发了一种灵敏且特异的巢式PCR检测方法用于检测粒细胞埃立克体。该检测方法扩增16S rRNA基因，并用于检测从7名临床表现符合人类粒细胞埃立克体病的患者身上采集的急性期乙二胺四乙酸抗凝血和血清样本。与仅鉴定出1份阳性样本的血清学检测方法相比，7例疑似病例中有5例采用全血提取的DNA作为模板通过PCR检测呈阳性。以相应血清样本提取的DNA作为模板的PCR检测鉴定出3份阳性样本。检测了该检测方法对人类样本的敏感性，结果显示检测限低于2个16S rRNA基因拷贝。该检测方法应用于非人类样本，结果显示从罗德岛采集的肩突硬蜱提取的模板DNA以及从马里兰州白尾鹿采集的乙二胺四乙酸抗凝血样本中扩增出了产物。所有PCR产物均进行了测序，并鉴定为粒细胞埃立克体特异的产物。在从蜱和鹿血样本中扩增出的产物中检测到了一种推定的变异粒细胞埃立克体16S rRNA基因序列。
