膜芯片快速诊断HBV基因型和耐药性方法的建立

发布时间：2018-03-18 06:43

本文选题：乙肝病毒　切入点：反向斑点杂交　出处：《中国现代医学杂志》2015年03期 　论文类型：期刊论文

【摘要】：目的基于多重PCR和反向斑点杂交原理,建立快速检测乙肝病毒(HBV)基因型和耐药性的膜芯片方法,并对其临床应用价值进行探讨。方法根据HBV基因序列设计探针并制作膜芯片,用PCR-反向斑点杂交检测3种HBV标准菌株和38例HBV临床分离株的基因型和耐药性,检测结果与测序结果进行比较,分析方法的准确性和最低检出限。对深圳地区187例HBV阳性标本进行基因分型和耐药性检测,了解其临床应用价值。结果 38例HBV临床分离株和3株HBV标准株鉴定结果与测序法完全一致,3株非HBV病毒和11株非HBV标准菌株无阳性显色,该方法准确性达到100%,最低检出限为103 copies/ml。深圳地区187例HBV临床分离株以B型为主,占57.8%,C型占37.4%,未测出D型。187例HBV临床分离株的耐药突变率为26.2%,以rt L180M+rt M204V/I为主,占67.3%,其次为rt M204I,占16.3%。临床HBV耐药株以C型为主,占65.3%,B型占34.7%。结论用膜芯片检测HBV基因型与耐药性具有简便、快速、准确性及特异性高的优点,评价指标符合临床实验要求,可在临床推广使用。
[Abstract]:Objective based on the principle of multiplex PCR and reverse dot blot hybridization, a method for rapid detection of HBV genotypes and drug resistance was established, and its clinical application value was discussed. Methods the probe was designed and the microarray was made according to the sequence of HBV gene. The genotypes and drug resistance of 3 HBV standard strains and 38 clinical isolates of HBV were detected by PCR- reverse dot blot. The results were compared with those of sequencing. The accuracy and minimum detection limit of the method were analyzed. The genotyping and drug resistance of 187 HBV positive samples from Shenzhen area were detected. Results the identification results of 38 HBV clinical isolates and 3 HBV standard strains were completely consistent with the sequencing method. No positive staining was found in 3 non-#en2# virus strains and 11 non-#en3# standard strains. The accuracy of this method was 100. The lowest detection limit was 103copias / ml. In Shenzhen area, 187 clinical isolates of HBV were mainly type B, accounting for 57.8% and C type, and the mutation rate of drug resistance in 187 clinical isolates of type D was 26.20.The main type was RT L180M rt M204V / I. The clinical HBV resistant strains were mainly type C, accounting for 65.3% of B type and 34.70.Conclusion Detection of HBV genotypes and drug resistance by membrane microarray is simple, rapid, accurate and specific, and the evaluation indexes meet the requirements of clinical experiments. It can be widely used in clinic.
【作者单位】：深圳市人民医院;
【基金】：深圳市科技工贸和信息化委员会基金资助项目(No:201103360)
【分类号】：R512.62

【参考文献】