Laboratory maintenance of Coxiella burnetii.
Laboratory maintenance of Coxiella burnetii.
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DOI：
10.1002/9780471729259.mc06c01s15
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发表时间：
2009-11-01
影响因子：
--
通讯作者：
Hendrix, Laura R
中科院分区：
文献类型：
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作者：
Samuel, James E;Hendrix, Laura R
伯纳蒂克希菌是一种专性细胞内革兰氏阴性细菌，是Q热的病原体，Q热是一种自限性流感样疾病，也可能表现为慢性心内膜炎。在气溶胶中以低感染剂量在环境中持续存在的能力导致伯纳氏梭菌被分类为BSL-3选择剂。常规繁殖是在胚胎卵或组织培养。从宿主组织中提纯包括多次差速离心，以从宿主物质中分离细菌。通过Gimenez染色、荧光染料或免疫荧光抗体（IFA）染色，在显微镜下常规验证宿主细胞的感染。采用PCR方法鉴定寄主材料中的伯氏疏螺旋体DNA。通过将光密度转换为干重，或者更准确地说，通过RT-PCR来确定基因组当量，可以完成纯化伯氏梭菌的定量。血清抗体滴度测定布氏疏螺旋体微凝集试验或ELISA。
Coxiella burnetii, an obligate intracellular Gram-negative bacterium, is the agent of Q fever, a self-limited flu-like illness that may also present as chronic endocarditis. The ability to persist in the environment at a low infectious dose in aerosols resulted in the classification of C. burnetii as a BSL-3 select agent. Routine propagation of this agent is in embryonated eggs or tissue culture. Purification from host tissues includes multiple differential centrifugations to separate bacteria from host material. Infection of host cells is routinely verified microscopically by using Gimenez stain, fluorescent dyes, or immunofluorescence antibody (IFA) staining. Identification of C. burnetii DNA in host material is measured by PCR. Quantification of purified C. burnetii is accomplished through conversion of optical density to dry weight or, more precisely, by RT-PCR to determine genome equivalents. Serum antibody titer to C. burnetii is determined by microagglutination assay or ELISA.