溶藻弧菌的依赖于核酸序列恒温扩增检测方法的建立

Establishment of the Nucleic Acid Sequence-based Amplification Method for Detecting Vibio Alginolyticus

采用自行建立和优化的依赖于核酸序列恒温扩增(NASBA)检测体系,对溶藻弧菌进行检测。采用溶藻弧菌的hsp60基因为目的片段设计特异性引物,建成可快速检测溶藻弧菌的NASBA检测法,并进行了特异性和灵敏度试验。结果表明:所建立起的NASBA检测方法,灵敏度为6.9×102 cfu.mL-1,高于普通PCR方法。溶藻弧菌的依赖于核酸序列恒温扩增检测方法具有较高灵敏度和和良好特异性,并且对仪器要求更低,用普通恒温设备即可进行反应,具有广阔的推广前景。

A new method, based on Nucleic Acid Sequence-based Amplification（NAS- BA） to detect Vibio alginolyticus of samples, was established. A highly specific set of primers was synthesized to target the hsp60 gene of Vibio algino[yticus so as to estah- lish Nucle tested. Th was highe was more ment. ic Acid Sequence-based Amplification method. Specificity and sensitivity were e results showed that the sensitivity of NASBA was 6.9X 102cfu ~ mL-~ which r than the result of PCR method. Detecting Vibio alginolyticus with NASBA specific and sensitive than PCR method and has lower instrumental require- ,there is a broad prospect.