登革病毒悬液芯片分型检测方法的建立  

Establishment of a multiplex liquid bead array for detecting and typing of dengue virus

作　　者：史玲莉[1] 闫冀焕[1] 李云[1] 罗明[2] 沈军[1] 韩伟[3] 

机构地区：[1]石家庄河北国际旅行卫生保健中心,050000 [2]北京市疾病预防控制中心 [3]河北出入境检验检疫局

出　　处：《国际病毒学杂志》2016年第3期173-176,共4页International Journal of Virology

基　　金：国家质检总局科研基金项目（2012IK225）

摘　　要：目的建立一种基于悬液芯片的登革病毒（dengue virus，DV）检测方法，可对四种血清型登革病毒进行快速检测和鉴定。方法依据GenBank上4种病毒的基因序列信息，设计并合成相关引物及探针序列。抽提病毒RNA，经反转录后对目的基因进行PCR扩增，产物与核酸探针微球组杂交后于Bio-Plex^TM 200系统检测荧光信号值。结果DV1的悬液芯片检测敏感性约9DNA拷贝，DV2、DV3、DV4的悬液芯片检测敏感性约90DNA拷贝。进而将本方法用于检测15份临床标本，其检测结果与分型荧光RT-PCR一致。结论建立了可同时检测四种血清型登革病毒的悬液芯片检测方法，为快速筛查和鉴定登革病毒提供了新的手段。Objective To establish a method for rapid detecting and genotyping of four serotypes of dengue virus （DV） based on a liquid bead array. Methods Primers and probes were designed and synthesized according to genomic sequences in Genbank. Viral RNA was extracted and reverse-transcribed, and target sequences were amplified using multiplex PCR. PCR products were hybridized with beads coupled with nucleic acid probes, and the fluorescence signals were detected by the Bio-PlexTM 200 system. Results The limit of detection for DV1 was about 9 DNA copies, and about 90 DNA copies for DV2, DV3, DV4. Fifteen clinical samples were detected using this method and 9 samples were positive, which was consistent with the detection results of real-time RT-PCR. Conclusions A liquid bead array assay of simultaneous detection for four serotypes of dengue viruses was developed, which will provide a new method for rapid screening and identification of dengue virus.

关 键 词：登革病毒 悬液芯片 多重PCR 

分 类 号：R440[医药卫生—诊断学]