Real-time RT-PCR Assay for the Detection of Culex flavivirus

作     者：CAO Yu Xi HE Xiao Xia FU Shi Hong HE Ying LI Hao GAO Xiao Yan LIANG Guo Dong WANG Huan Yu 

作者机构：State Key Laboratory of Infectious Disease Prevention and Control(SKLID)Department of Viral EncephalitisNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and Prevention Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases 

出 版 物：《Biomedical and Environmental Sciences》 (生物医学与环境科学（英文版）)

年 卷 期：2015年第28卷第12期

页      面：917-919页

核心收录：

学科分类：100208[医学-临床检验诊断学] 1004[医学-公共卫生与预防医学(可授医学、理学学位)] 1002[医学-临床医学] 100401[医学-流行病与卫生统计学] 10[医学] 

基　　金：supported by grants from the Development Grant of State Key Laboratory of Infectious Disease Prevention and Control(2012SKLID204,2015SKLID505) the Ministry of Science and Technology of People’s Republic of China(No.2013ZX10004101) 

主　　题：PCR Real-time RT-PCR Assay for the Detection of Culex flavivirus RT time 

摘      要：Based on the Culex flavivirus (CxFV) E gene sequences in GenBank, CxFV-specific primers and probes were designed for real-time reverse transcription-polymerase chain reaction (RT-qPCR). The specificity test revealed that CxFV could be detected using RT-qPCR with the specific CxFV primers and probes; other species of arboviruses were not detected. The stability test demonstrated a coefficient of variation of 1.5%. A quantitative standard curve for CxFV RT-qPCR was established. Quantitative standard curve analysis revealed that the lower detection limit of the RT-qPCR system is 100 copies/mu L. Moreover, RT-qPCR was used to detect CxFV viral RNA in mosquito pool samples. In conclusion, we established a real-time RT-PCR assay for CxFV detection, and this assay is more sensitive and efficient than general RT-PCR. This technology may be used to monitor changes in the environmental virus levels.