HCV基因型检测芯片的设计与研制初报  

A pilot study on design and develop of biochip for HCV genotyping

作　　者：齐名[1] 甄国亮[2] 熊华[1] 刘新钰[3] 吴引伟[3] 袁春伟[2] 武建国[1] 

机构地区：[1]南京军区南京总医院,南京210002 [2]东南大学生物医学工程系 [3]南京市第二医院

出　　处：《临床检验杂志》2002年第6期333-336,共4页Chinese Journal of Clinical Laboratory Science

基　　金：江苏省社会发展科技基金 (BS2 0 0 0 0 2 5 )

摘　　要：目的　设计与研制丙型肝炎病毒 (HCV)基因型检测芯片。方法　用生物信息学方法设计HCV 5′UTR型特异性探针。按设计模式 ,以自动化微阵列点样仪将探针点样于APTES PDC修饰的玻片表面。用商品PCR试剂盒掺入荧光分子对患者血清样本进行扩增 ,用激光共聚焦荧光扫描仪检测并分析结果。结果　根据 5′UTR序列设计了各型HCV的特异性探针。芯片表面修饰符合固定探针和杂交检测的需求。含荧光分子的RT PCR产物可成功地与固定于芯片上的探针阵列杂交进行HCV基因型的检测。 2 1例丙型肝炎患者中 ,19例为 1b型 ,1例为 2a型 ,1例为混和型。结论　所设计研制的基因芯片可望成为检测HCV基因型的操作简便 ,结果准确 ,价格低 。Objective To design and develop a biochip for genotyping of Hepatitis C virus.Methods The type-specific probes for HCV 5′UTR were designed according to bioinformatics principles. The probes were spotted on the surface of APTES-PDC-modified glass slides by microarray spotter tracing the designed patterns. The RNA sequences in sera sample from patients with hepatitis C were amplified by using commercial RT-PCR kit with incorporate fluorescent molecules and the slides were determined by Laser cofocus fluorescent scanner and the results were analysed by designed computer program.Results The probes for 5′UTR of HCV of each type were designed. The modified surface of slides met with the requirement for probes coupling and hybridization. The products of fluorescent RT-PCR were successfully hybridized with the probes immobilized on designed biochip to genotype HCV in samples.Among 21 patients with hepatitis C 19 were type 1b, 1 was 2a and 1 was mixed 1b and 2a.Conclusions The biochip which was designed and developed in this study as a new experimental tool is apt to determine HCV genotyping with simple operation, accurate results, low price and less time-consuming.

关 键 词：丙型肝炎病毒 基因型 生物芯片 设计 研制 

分 类 号：R512.6[医药卫生—内科学]