关键词: 白纹伊蚊, 登革-2型病毒, TaqMan MGB探针, 实时聚合酶链反应

Abstract: Objective To study the sensitivity of TaqMan MGB Real-time Polymerase Chain Reaction (PCR) on detecting Dengue virus (DV) from Aedes albopictus and to develop a sensitive,specific,and repetitive assay method for DV diagnosis. Methods Females A.albopictus were artificially infected with Den-2 in lab,different concentration for infecting mosquito (mosquitos/ 500 μl or mosquitoes/ 1 000 μl) and different pools (0,10,25,50 females of lab A.albopictus without DV were added to each treatment) were designed to process of virus detection by one-step and two-step TaqMan MGB Real-time PCR. Fluorescent signal was observed to show the results. Results For all pools,the minimal concentration which could be detected was 2 mosquitoes/ 500 μl and 3 mosquitoes/ 1 000 μl for two-step TaqMan MGB Real-time PCR,but 5 mosquitoes/ 500 μl for one-step TaqMan MGB Real-time PCR. There was no evident effect on the detection of DV RNA by the RNA of A.albopictus. Conclusion TaqMan MGB is sensitive and specific to detect DV,and is a perfect surveillance method for DV index of A.albopictus. It is better for 20-30 mosquitoes/pool and 500 μl dilution/pool by two-step TaqMan MGB Real-time PCR.