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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Evaluation of PCR-Based Assay for Diagnosis of Spotted Fever Group Rickettsiosis in Human Serum Samples
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Evaluation of PCR-Based Assay for Diagnosis of Spotted Fever Group Rickettsiosis in Human Serum Samples

机译：基于PCR的人血清样品中发现的发烧组Ri病的诊断方法的评估

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A nested PCR assay was developed for the detection of spotted fever group (SFG) rickettsiae in serum samples. The assay was based on specific primers derived from the rickettsial outer membrane protein B gene (rompB) of Rickettsia conorii. An SFG rickettsia-specific signal is obtained from R. akari, R. japonica, R. sibirica, and R. conorii. Other bacterial species tested did not generate any signal, attesting to the specificity of the assay. As few as seven copies of the rompB gene of R. conorii could be detected in 200 μl of serum sample. The assay was evaluated with a panel of sera obtained from patients with acute-phase febrile disease tested by immunofluorescent antibody assay (IFA). The SFG rickettsia-specific DNA fragment was detected in 71 out of 100 sera, which were proven to have immunoglobulin M antibodies against SFG rickettsial antigen by IFA. The results were further confirmed by restriction fragment length polymorphism and sequencing analysis of the DNA fragments. The results indicated that this PCR assay is suitable for the diagnosis of spotted fever group rickettsiosis in Korea.
机译：开发了巢式PCR检测试剂盒，用于检测血清样品中的点状发热组（SFG）立克次体。该测定基于源自 Rickettsia conorii 的立克次氏菌外膜蛋白B基因（ rompB ）的特异性引物。 SFG立克次体特异性信号获自 R。 akari ， R。 japonica ， R。 sibirica 和 R。 conorii 。测试的其他细菌种类未产生任何信号，证明了测定的特异性。 R的 rompB 基因少至七个副本。在200μl血清样品中可以检测到conorii 。用从免疫荧光抗体测定法（IFA）测试的急性期发热性疾病患者的血清中评估该测定法。在100份血清中的71份中检测到SFG立克次体特异性DNA片段，该片段经IFA证实具有针对SFG立克次体抗原的免疫球蛋白M抗体。通过限制性片段长度多态性和DNA片段的测序分析进一步证实了该结果。结果表明，该PCR检测方法适用于韩国斑疹状立克次氏病的诊断。

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