中国动物传染病学报2024，Vol.32Issue(6)：63-68,6.
对虾白斑综合症病毒LAMP-TaqMan检测方法的建立及应用
Development and Application of Lamp-TaqMan Detection Method for White Spot Syndrome Virus
摘要
Abstract
In order to improve the specificity of loop-mediated isothermal amplification(LAMP)detection method,this study developed a LAMP-TaqMan method to detect White spot syndrome virus(WSSV)nucleic acid in shrimps.In this study,the test plasmid was constructed based on the conserved sequences of the WSSV genome and LAMP primer sets were designed and screened accordingly.The loop primer modified by the fluorescent quenching group was used as the TaqMan probe in the LAMP-TaqMan detection method.The results show that the LAMP-TaqMan method has good specificity in the detection of WSSV.The optimal detection temperature was 63℃and the linear correlation coefficient R2 of the test plasmid was 0.990.The minimum detection limit was 0.76×101 copies/µL with good repeatability.The coefficient of variation CV values of the results were all less than 3%with good reproducibility.Total 60 shrimp samples were tested in parallel by this LAMP-TaqMan method and a commercial detection kit.As a result,the LAMP-TaqMan method had accuracy at 96.67%and sensitivity at 100.00%.Taken together,the LAMP-TaqMan constant temperature nucleic acid method had the accurate and rapid advantages with low detection limit and effectively avoided the"false positive"results caused by non-specific amplification of LAMP.Therefore,the LAMP-TaqMan method had a wide market prospect in the accurate and rapid detection of aquatic diseases.关键词
LAMP/Taqman探针/对虾白斑综合症病毒/核酸检测Key words
LAMP/TaqMan probe/White spot syndrome virus/nucleic acid detection分类
农业科技引用本文复制引用
刘昱,柳梦思,董振国,焦义然,时国强,石磊..对虾白斑综合症病毒LAMP-TaqMan检测方法的建立及应用[J].中国动物传染病学报,2024,32(6):63-68,6.基金项目
河北省产业创新创业团队项目(205A7601D) （205A7601D）
