基于LAMP技术建立解脲支原体基因试纸条检测方法

目的 建立一种灵敏度高、特异性强、检测速度快的方法检测解脲支原体.方法 基于环介导恒温扩增技术(LAMP),根据解脲支原体序列特征设计3对引物进行解脲支原体DNA切口酶核酸恒温扩增,扩增过程在一对引物中标记生物素,随着扩增的进行生物素直接引入扩增片段中,扩增结束后产物在密闭装置中进行免疫试纸条显色反应,根据显色卡的颜色判定结果的阴阳性.结果 该技术检测解脲支原体较实时荧光PCR技术灵敏度要高10倍以上,其它病原体检测均阴性该方法特异性与实时荧光PCR技术相当.结论 恒温扩增联合试纸条技术检测解脲支原体具有较高的敏感性和特异性,检测速度快,适合各医院开展.

Detection of Mycoplasma Urealytium Gene with Test Strip Based on LAMP

Objective Establishment of a detection method for urea mycoplasma with high sensitivity,high specificity,and high speed.Methods Based on LAMP,three pairs of Primers were designed to amplify the urea mycoplasma sequences at constant temperature.One pair of primers was marked by biotin,and the product was detected by immune dipsticks color reaction in the progress of amplification in an airtight device.The results were analyzed based on the changing of color in samples.Results Successfully established the LAMP test strip methods for urea mycoplasma detection.The sensitivity of this methods is more than ten times higher than the that of real-time PCR.AS other pathogenic tests were negative,the specificity of this method is similar to that of the real-time PCR.Conclusion The isothermal amplification associated with dipstick technology showed high sensitivity and specificity.Therefore,this method may be suitable for all hospitals.