甲型H1N1流感病毒HA基因RT-LAMP检测方法的建立与初步应用

摘要

Objective: To establish the methods of RT-LAMP assay in influenza virus A(H1N1) HA gene and use to detect clinical samples. Methods: RT-LAMP primer for HA of H1N1 was designed by using Primer Explorer V4 on line, and the sensitivity and specificity of the established assay was evaluated. Results: Comparing with traditional RT-PCR, RT-LAMP showed fine specificity and higher sensitivity until 10 copy. By RT-LAMP assay we got 2 positive samples in 76 clinical swabs from the children with respiratory infections. Conclusion: RT-LAMP assay for detection of H1N1 is sensitive, efficient, rapid and simple. These results imply potential on-the-spot detection of influenza H1N1 by RT-LAMP method.%目的:建立反转录环介导等温扩增技术(RT-LAMP)检测甲型H1N1流感病毒HA基因的方法,并用于检测临床样本.方法:通过在线软件Primer Explorer V4设计H1N1 HA基因的RT-LAMP引物,建立RT-LAMP检测方法,并评价其灵敏度和特异度.结果:与传统RT-PCR方法相比,RT-LAMP检测方法具备更高的灵敏度,达到10个拷贝,并且具有良好的特异性；在76份呼吸道感染儿童的咽拭子标本中检测到2份阳性,与RT-PCR方法检测结果相同.结论:建立的H1N1 RT-LAMP检测方法灵敏特异,简单快速,具备H1N1现场检测应用前景.