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 108 copies/μL to 102 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.

中文翻译：

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用于检测新兴传染性伊丽莎白女王的实时荧光定量PCR检测方法。

革兰氏阴性芽孢杆菌伊丽莎白女王微藻正以威胁人类生命的病原体的形式出现。但是，不存在检测米氏大肠杆菌的特异性和快速的诊断方法。在这里，我们建立了一种实时PCR检测试剂盒，用于快速，灵敏和特异地检测米氏大肠杆菌，其动态范围很宽，为108拷贝/μL至102拷贝/μL。实时测定的检出限为145拷贝/μL，比常规PCR灵敏度高100倍。来自不同宿主物种的所有临床分离株大肠埃希菌均产生非常接近的Tm（80.25±0.25°C）。另外，在非靶细菌物种的测定中未观察到交叉反应或假阳性。该测定的性能主要通过测试青蛙组织样品来评估。总体而言，我们的研究提供了一种实时PCR检测方法，该方法快速，