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首页> 外文期刊>Journal of Clinical Microbiology >Evaluation of Coxiella burnetii Antibiotic Susceptibilities by Real-Time PCR Assay
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Evaluation of Coxiella burnetii Antibiotic Susceptibilities by Real-Time PCR Assay

机译：实时荧光定量PCR法测定伯氏柯氏杆菌的抗生素敏感性

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Coxiella burnetii is an obligate intracellular bacterium. The inability to cultivate this organism on axenic medium has made calculation of infectious units challenging and prevents the use of conventional antibiotic susceptibility assays. A rapid and reliable real-time PCR assay was developed to quantify C. burnetii cells from J774.16 mouse macrophage cells and was applied to antibiotic susceptibility testing of C. burnetii Nine Mile, phase I. For calculation of bacterial replication, real-time PCR performed equally as well as immunofluorescent-antibody (IFA) assay when J774.16 cells were infected with 10-fold serial dilutions of C. burnetii and was significantly (P < 0.05) more repeatable than IFA when 2-fold dilutions were used. Newly infected murine macrophage-like J774.16 cells were treated with 8 μg of chloramphenicol per ml, 4 μg of tetracycline per ml, 4 μg of rifampin per ml, 4 μg of ampicillin per ml, or 1 μg of ciprofloxacin per ml. After 6 days of treatment, tetracycline, rifampin, and ampicillin significantly (P < 0.01) inhibited the replication of C. burnetii, while chloramphenicol and ciprofloxacin did not. In general, these results are consistent with those from prior reports on the efficacy of these antibiotics against C. burnetii Nine Mile, phase I, and indicate that a real-time PCR-based assay is an appropriate alternative to the present methodology for evaluation of the antibiotic susceptibilities of C. burnetii.
机译： burnetii 是专性的细胞内细菌。由于无法在抗微生物培养基上培养这种生物，因此感染单位的计算变得具有挑战性，并阻止了常规抗生素敏感性试验的使用。开发了一种快速可靠的实时PCR分析方法来定量C。 J774.16小鼠巨噬细胞产生的Burnetii 细胞，并用于 C的抗生素敏感性测试。九英里，第一阶段。为了计算细菌复制，当J774.16细胞被10倍系列稀释的感染时，实时PCR和免疫荧光抗体（IFA）分析同样有效。 C。当使用2倍稀释液时，Burnetii 的可重复性（ P <0.05）比IFA明显得多。用每毫升8μg氯霉素，每毫升4μg四环素，每毫升4μg利福平，每毫升4μg氨苄西林或每毫升1μg环丙沙星处理新感染的鼠类巨噬细胞样J774.16细胞。治疗6天后，四环素，利福平和氨苄青霉素显着（ P <0.01）抑制了 C的复制。 Burnetii ，而氯霉素和环丙沙星则没有。通常，这些结果与先前关于这些抗生素对 C的功效的报道一致。 Burnetii 九英里区（I），表明基于实时PCR的检测方法是本方法评估 C抗生素敏感性的适当方法。 Burnetii 。

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