Research Virology Department, State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise, Kyiv, Ukraine.
State Scientific Control Institute of Biotechnology and Strains of Microorganisms, Kyiv, Ukraine.
Vector Borne Zoonotic Dis. 2020 Feb;20(2):100-106. doi: 10.1089/vbz.2019.2518. Epub 2019 Sep 19.
is an obligate intracellular pathogen and the causative agent of Q fever. In Ukraine, 28 human cases of Q fever were reported between 1997 and 2006; however, there are no state-approved, standardized molecular diagnostic assays that can be used systematically to investigate transmission to humans and its distribution throughout Ukraine. To address this deficiency, we followed the recommendation of the World Organization for Animal Health (OIE) and developed a confirmatory PCR for for veterinary diagnosis in Ukraine. The PCR assay targeted the outer membrane-associated gene com1 in . Oligonucleotide primers were selected that amplify a 689-bp DNA fragment of the gene (primers: CoxF2 = 5'-ACYGCAGGCGTGGCGATAG-3' and CoxR4 = 5'-TGAAGGTTTTGTTGTGAGGTGGC-3'). The assay proved highly sensitive and specific to DNA detection (LOD = 0.37 pg/μL). Reproducibility of the test was verified by comparing the PCR results with those of a different PCR protocol and qPCR. Using the CoxF2/CoxR4 primer set and reaction conditions described here, the PCR Diagnostic Kit PCR-TEST was developed and officially registered for use in Ukraine by the State Scientific Control Institute of Biotechnology and Strains (Kyiv, Ukraine) for diagnostic purposes.
是一种专性细胞内病原体，也是 Q 热的病原体。在乌克兰，1997 年至 2006 年间报告了 28 例人类 Q 热病例；然而，没有经过国家批准的标准化分子诊断检测方法可以系统地用于调查其向人类的传播及其在乌克兰的分布。为了解决这一不足，我们遵循世界动物卫生组织（OIE）的建议，为兽医诊断开发了一种用于的确认性 PCR。PCR 检测针对的是外膜相关基因 com1 。选择了扩增基因（引物：CoxF2=5'-ACYGCAGGCGTGGCGATAG-3'和 CoxR4=5'-TGAAGGTTTTGTTGTGAGGTGGC-3'）的 689bp DNA 片段的寡核苷酸引物。该检测方法对 DNA 的检测具有高度的敏感性和特异性（LOD=0.37pg/μL）。通过将 PCR 结果与另一种 PCR 方案和 qPCR 进行比较，验证了该试验的重现性。使用 CoxF2/CoxR4 引物组和本文描述的反应条件，开发了 PCR-TEST 诊断试剂盒，并由乌克兰国家生物技术和品系科学控制研究所（基辅，乌克兰）正式注册，用于诊断目的。
