龟致病性维氏气单胞菌和鲍曼不动杆菌双重PCR检测方法的建立

Establishment of Duplex PCR for Detection of Aeromonas veronii and Acinetobacter baumannii from Turtles

应用2对特异性引物,通过对反应条件和体系的优化,成功建立了能快速检测维氏气单胞菌和鲍曼不动杆菌两种病原的双重PCR方法。研究结果表明,该方法特异性强,仅对龟鳖维氏气单胞菌和鲍曼不动杆菌有扩增,而对其他龟鳖常见病原无扩增;该方法对维氏气单胞菌最低检测核酸质量浓度为1.75×10-3 ng/μL,对鲍曼不动杆菌最低检测核酸质量浓度为3.96×10-3 ng/μL,检出率较传统检测方法高。该研究所建立的双重PCR方法灵敏度高、特异性好,可为临床龟鳖维氏气单胞菌和鲍曼不动杆菌感染的诊断和流行病学调查等提供帮助。

In this study,a duplex PCR（dPCR）was established and optimized to detect pathogenic bacteria Aeromonas veronii and Acinetobacter baumannii simultaneously in turtles,and two sets of specific primers were designed for development of the duplex PCR.After optimization of PCR components and reaction profile,a duplex PCR method was established.The results showed that the assay is highly specific to A.veronii and A.baumannii,and no-target pathogens were detected with the minimum detectable nucleic acid concentration of 1.75×10-3 ng/μL in A.veronii and 3.96×10-3 ng/μL in A.baumannii,more sensitive than conventional methods.The duplex PCR established in this study is specific and sensitive,and will be useful for clinical detection and identification of A.veronii and A.baumannii.