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-PlexTM 200系统检测荧光信号值.结果 DV1的悬液芯片检测敏感性约9 DNA拷贝,DV2、DV3、DV4的悬液芯片检测敏感性约90 DNA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.

abstractsObjective 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.