链置换恒温扩增快速检测单核细胞增生李斯特氏菌

Rapid detection of Listeria monocytogenes by strand displacement amplification

中文关键词:  单核细胞增生李斯特氏菌  链置换恒温扩增

英文关键词:Listeria monocytogenes  strand displacement amplification

中文摘要:

      目的 开发单核细胞增生李斯特氏菌快速、简便、准确的链置换恒温扩增(strand displacement amplification, SDA)检测方法。方法 根据单核增生李斯特菌特异性基因prfA基因片段, 设计链置换扩增特异性引物, 建立单核细胞增生李斯特菌SDA快速检测方法, 同时对设计的引物的特异性和灵敏度进行研究。 结果 所建立的单核细胞增生李斯特菌SDA扩增方法灵敏度达到了7.0×102 CFU/mL, 检验技术可以特异性地扩增单核增生李斯特菌。结论 本试验所建立的单核细胞增生李斯特菌SDA检验技术具有快速、简便、特异性强的特点, 具有在进出口实验室推广应用前景。

英文摘要:

      Objective To develop a rapid, simple and accurate method for detection of Listeria monocytogenes by strand displacement amplification (SDA). Methods Chain replacement amplification primers were designed according to the sequence of prfA genetic fragment of Listeria monocytogenes. A rapid detection method for Listeria monocytogenes was established by SDA. The specificity and sensitivity of the designed primers were investigated. Results The sensitivity of the established method for SDA amplification of Listeria monocytogenes was 7×102 CFU/mL. It showed that the SDA technology could specially amplify Listeria monocytogenes. Conclusion The established SDA detection technique of Listeria monocytogenes is rapid, simple and specific, which has the prospect of popularization and application in the port.