环介导技术在细菌性痢疾检测中的应用

摘要

Objective:To establish and apply the methods of LAMP assay for the detection of Shigella of infectious diarrhea patients from the enteric clinicsin. Methods:The specific primers of the conserved regions of Shigella by using Primer Explorer V5 online and composed to establish the LAMP assay for detection of Sal-monella. The 760 fecal specimens among diarrhea patients visiting the enteric clinicsin in Hospital of Chengde Medical College from 2016 was collected and test by Real-time PCR ,LAMP and the Traditional Method. The specificity and sensibility of LAMP were evaluated by comparing with Real-time PCR and the Traditional. Re-sults:There was no statistical difference in the specificity of 3 method( P>0.05) . LAMP showed fine specifici-ty and high sensitivity. Conclusion:LAMP assay was performed in water oven and the results could be directly determined by eyes under visible light or UV lamp. Because of its rapidity and minimal equipment require-ment,LAMP assay for detection of Shigella in primary diseases control and prevention agency and primary health institutions.%目的:建立环介导等温扩增法(LAMP)检测肠道门诊感染性腹泻病例志贺菌.方法:应用Primer Explorer V5软件设计针对志贺菌的保守区引物,建立LAMP检测方法,采用荧光定量PCR法、LAMP法和分离培养法三种方法,对760例承德医学院附属医院肠道门诊腹泻患者粪便标本进行检测.对建立的方法进行特异度、灵敏度试验,与荧光定量PCR、培养法进行比对.结果:3种方法的检测结果差异无统计学意义(P>0.05).LAMP检测方法具有良好的敏感度和特异度.结论:LAMP法使用水浴锅进行等温扩增,结果可在可见光或紫外灯下直接肉眼观察.LAMP检测方法灵敏特异,简单快速,无需特殊仪器,可广泛用于基层疾控医疗机构志贺菌的检测.