基于白纹伊蚊Actin基因参照的登革Ⅱ型病毒一步法实时荧光定量PCR检测方法的建立  

ESTABLISHMENT OF ONE-STEP REAL-TIME FLUORESCENT QUANTITATIVE PCR METHOD FOR DETECTION OF DENGUE VIRUS TYPE Ⅱ BASED ON THE ACTIN GENE OF AEDES ALBOPICTUS

作　　者：李超杰 姜玉庭 张强辉 高剑 刘源[1] 邢丹 李春晓 张恒端 郭晓霞 赵彤言 LI Chao-Jie;JIANG Yu-Ting;ZHANG Qiang-Hui;GAO Jian;LIU Yuan;XING Dan;LI Chun-Xiao;ZHANG Heng-Duan;GUO Xiao-Xia;ZHAO Tong-Yan(Beijing Key Laboratory of Vector Borne and Natural Focus Infectious Diseases,State Key Laboratory of Pathogen and Biosecurity,Institute of Microbiology and Epidemiology,AMM5,Beijing 100071,China)

机构地区：[1]军事科学院军事医学研究院微生物流行病研究所,病原微生物生物安全国家重点实验室,媒介生物危害和自然疫源性疾病北京市重点实验室,北京100071

出　　处：《寄生虫与医学昆虫学报》2020年第4期239-243,共5页Acta Parasitologica et Medica Entomologica Sinica

基　　金：“十三五”传染病重大专项(No.2017ZX10303404)。

摘　　要：本研究旨在建立一种以白纹伊蚊Actin基因表达量作为参照的登革Ⅱ型病毒一步法实时荧光定量PCR检测技术,为登革病毒在蚊虫中的定量检测提供更准确的检测方法。根据登革Ⅱ型病毒和白纹伊蚊Actin基因保守区序列设计引物和探针,在检测体系中同时加入病毒和Actin基因的特异性引物,计算每个样本中病毒含量与Actin表达量的比值,得到更为准确的定量结果。该方法重复性好、灵敏度高,可用于实验室白纹伊蚊感染登革Ⅱ型病毒的病毒载量检测。To provide a more accurate method for detecting dengue virus from mosquitoes,an one-step real-time fluorescent quantitative PCR method was established taking the expression of Actin gene of Aedes albopictus as reference.The primers and probes were designed based on the dengue type 2 virus and the conserved region of Actin gene of Ae.albopictus,and then added into the detection system at the same time.The ratio of virus content to Actin expression in each sample was calculated to get more accurate quantitative results.The technique has good repeatability and high sensitivity,and can be used to detect the dengue II viral load of Ae.albopictus infected in both laboratory and field.

关 键 词：登革Ⅱ型病毒 一步法实时荧光定量PCR 白纹伊蚊 ACTIN 

分 类 号：R384.1[医药卫生—医学寄生虫学]