摘要
目的评价基因芯片技术在HBV基因型及HBVP基因YMDD变异检测中的应用价值,并对不同基因型病例的临床情况进行分析。方法抽提HBV DNA,用PCR方法扩增S基因及P基因部分区域,扩增产物与基因芯片上的寡核苷酸探针进行杂交,杂交结果通过芯片影像读取仪扫描到计算机上,经软件分析可得到HBV基因型、HBVYMDD野生型及YVDD、YIDD变异型结果;部分标本通过HBV preS/S基因序列测序证实。结果38份高病毒滴度标本(HBV DNA定量>1.0×105拷贝/毫升)中22例为B基因型(58%),16例为C基因型(42%);32例为HBV YMDD野生型,2例为YVDD变异型、4例为YIDD变异型。临床情况方面,无症状携带者中有2例为基因型B、3例为基因型C,慢乙肝患者中有16例为基因型B1、2例为基因型C,肝硬化患者中有4例为基因型B、1例为基因型C;B型和C型患者e抗原阳性率分别为54.5%(12/22)和87.5%(14/16)。10份HBV DNA定量<1.0×105拷贝/毫升的标本不能检测出HBV基因型及YMDD变异情况。11例阳性标本的基因测序结果与基因芯片检测结果一致。结论(1)基因芯片检测HBV基因型及HBV P基因区YMDD变异准确性高、特异性好,同时每次实验得到的信息量多,适合于临床开展应用,但需提高灵敏度;(2)C基因型乙肝患者HBV e抗原阳性率高于B型患者。
Objective To evaluate the application value of gene chip technology in the detection of Hepatitis B Virus genotypes and p gene YMDD mutations, and analyse the clinical status of different genotype cases. Methods The serum samples from 48 patients infected with HBV were detected by gene chip technique for HBV genotypes,YMDD wildtype and YVD or YIDD mutations, some positive samples had been testified by HBV preS/S gene sequencing.Results In 38 samples which HBV DNA quantifies were higher than 105 copies/ml,22 eases were genotype B,16 cases were genotype C;32 cases were YMDD wildtype,2 cases were YVDD mutation,4 eases were YIDD mutation.In the clinical status,2 eases were genotype B and 3 eases were genotype C in the absence of symptoms carriers, 16 eases were genotype B and 12 cases were genotype Cin the chronic hepatitis B patients,4 eases were genotype B and 1 eases were genotype C in the cirrhosis patients;the HBV e antigens were positive in 14/16 eases of genotype C and 12/22 cases of genotype B. 10 samples which HBV DNA quantities were lower than 105 eopies/ml couldn't be detected the HBV genotypes and YMDD mutations. DNA sequence analysis of 11 positive samples verified that gene chips were available. Conclusion (1) Gene chip technique for the detection of HBV genotypes and YMDD mutations is reliable and its specificity is high, it can give more information in one test and could be used for clinical application, but it is needed to improve the sensitivity; (2)The ttBeAg positive rate of genotype C is higher than that of genotype B.
出处
《中国实验诊断学》
2007年第2期207-212,共6页
Chinese Journal of Laboratory Diagnosis
基金
广州市卫生局重点课题(编号:2004Z008)资助。