病毒性肝炎联合诊断寡核苷酸芯片的临床应用性研究  

Clinical application of a oligo microarray assay for simultaneous detection of HBV,HCV and HDV

作　　者：孙朝晖[1] 危敏[2] 王蜀燕[1] 王从容[3] 石玉玲[1] 李林海[1] 

机构地区：[1]广州军区广州总医院检验科 [2]南方医科大学基因工程研究所,广州510515 [3]广州市南方医院检验科,广州510515

出　　处：《中国卫生检验杂志》2007年第10期1753-1756,共4页Chinese Journal of Health Laboratory Technology

基　　金：广东省自然科学基金(7000065);中国博士后科学基金(20070410831);广东省医学科研基金(A2007477)

摘　　要：目的:制备联合诊断乙型(HBV)、丙型(HCV)、丁型(HDV)肝炎病毒寡核苷酸(Oligo)芯片,并进行系列优化及临床应用性研究。方法:首先利用软件Array designer 3.0对BLAST检索所得的HBV、HCV、HDV种属保守序列逐一分析,设计出60mer Oligo探针。用PixSys5500型基因芯片打印仪将设计好的探针打印到氨基化玻片上制备成基因芯片,并对杂交条件进行系列优化以适应临床血清标本的检测。结果:芯片杂交条件进行优化后,芯片检测的敏感性、特异性都大大增强,还缩短了检测时间;杂交结果显示,Oligo芯片检测HBV、HCV、HDV的敏感性、特异性分别为92.2%,85.7%,91.3%和96.0%,94.5%,98.0%。结论:制备的长链Oligo集合肝炎病毒检测芯片检测敏感性、特异性均佳,为下一阶段进行中试规模的临床血清样品检测做好了准备,同时也为集合更多种、不同亚型肝炎病毒联合检测芯片的制备打下了坚实的基础。Objective：A model oligo microarray assay for the simultaneous detection of HBV,HCV and HDV were developed.Modified protocols were used and applied to clinical sample detection.Methods：Oligo probes were designed with the Array designer 3.0 program according to the conserved region of HBV,HBV and HDV after BLAST.The microarray was prepared by spotting oligo probes on the surface of amido-modified glass slide using the robotics system.In the present study,modified protocols were used and applied to clinical sample detection.Results：The diagnostic validity of the microarray method was evaluated after the modified protocols.The results of hybridization and sequence data analysis showed that the specificity and sensitivity in the detection of HBV and HCV were 92.2%,85.7%,91.3% and 96.0%,94.5%,98.0%,respectively.Conclusion：Using oligo probes to construct gene chips for simultaneous clinical diagnosis of hepatitis virus results in a rapid,simple and effective method.The model microarray assay may be widely used in the clinical diagnosis of hepatitis virus and genotypes.

关 键 词：肝炎病毒/乙型/丙型/丁型 寡核苷酸 探针 基因芯片 杂交 

分 类 号：R942[医药卫生—药剂学]