摘要
目的建立一种简便、快速的以DNA芯片技术为基础的丙型肝炎病毒基因分型检测方法。方法 根据丙型肝炎病毒HCV的基因序列信息设计分型探针,将采用荧光标记的寡核苷酸引物所扩增的丙型肝炎病毒基因产物与结合在芯片上的特异性探针进行快速杂交,通过扫描荧光强度值定性和定量判定结果。结果 应用本法对65例患者血清中的丙型肝炎病毒核酸进行基因分型,对部分标本检测结果与测序法进行对比,吻合率达100％,说明检测结果有很高的特异性。结论 我们所用DNA芯片技术检测HCV RNA及其基因分型,简便、快速、特异性好、灵敏度高、结果判定指标客观,可望在临床推广使用。
Objective To establish a new method based DNA chip technique for detecting HCV genotypes. Methods Genotyping probes were designed according to the sequence of HCV 5' NCR to generate DNA chip. The probes on DNA chip contains 5 major genotypes and 8 subtypes. The DNA fragment amplified by labeling CyS fluorescence was hybridized with DNA chip.Results Fifty-five out of 65 isolates detected by DNA chip belonged to 1b. DNA sequencing of form a part of the isolates was used as the control . The results of both were completely consistent. Conclusion The method is simple and rapid with high specificity and sensitivity . It can be applied in detection of HCV RNA and genotypes.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2003年第4期375-377,共3页
Chinese Journal of Experimental and Clinical Virology
基金
中科院重大项目(KSCX1-06)
上海市科发基金资助项目(99QD14030)
