Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China; Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention, Wuhan, 430070, China; ShuangshuishuiShuanglü Institute, Huazhong Agricultural University, Wuhan, 430070, China.
Department of Aquatic Animal Medicine, College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China; Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention, Wuhan, 430070, China; ShuangshuishuiShuanglü Institute, Huazhong Agricultural University, Wuhan, 430070, China.
Mol Cell Probes. 2020 Aug;52:101571. doi: 10.1016/j.mcp.2020.101571. Epub 2020 Apr 11.
Elizabethkingia miricola, a Gram-negative bacillus, is emerging as a life-threatening pathogen in both humans and animals. However, no specific and rapid diagnostic method exists to detect E. miricola. Here, we established a real-time PCR assay for the rapid, sensitive, and specific detection of E. miricola with a wide dynamic range of 10 copies/μL to 10 copies/μL. The detection limit of the real-time assay was 145 copies/μL, which was 100 times more sensitive than conventional PCR. All clinical isolates E. miricola from different host species yield very close Tm (80.25 ± 0.25 °C). Additionally, no cross-reaction or false positives were observed in the assay for non-target bacterial species. The performance of this assay was primarily assessed by testing frog tissue samples. Overall, our study provided a real-time PCR assay, which is a rapid, sensitive, and specific diagnostic method that could be used for early diagnosis and epidemiological investigation of E. miricola.
伊丽莎白菌，一种革兰氏阴性杆菌，正成为人类和动物健康的一种潜在威胁病原体。然而，目前还没有特定的、快速的诊断方法可以检测到伊丽莎白菌。在这里，我们建立了一种实时 PCR 检测方法，可以快速、灵敏、特异性地检测伊丽莎白菌，其动态范围为 10 拷贝/μL 至 10 拷贝/μL。实时检测方法的检测限为 145 拷贝/μL，比常规 PCR 灵敏 100 倍。所有来自不同宿主物种的临床分离的伊丽莎白菌都具有非常接近的 Tm（80.25±0.25°C）。此外，在检测非目标细菌种类时，没有观察到交叉反应或假阳性。该方法的性能主要通过检测蛙组织样本进行评估。总的来说，我们的研究提供了一种实时 PCR 检测方法，该方法快速、灵敏、特异性高，可用于伊丽莎白菌的早期诊断和流行病学调查。
