HBV基因芯片的制备及其在临床应用的研究
发布时间：2018-12-21 08:14
【摘要】：前言 乙肝病毒(HBV)的感染呈世界性的分布,乙肝病毒基因组分为多个亚型,一种是根据S区的氨基酸差异划分的血清型,一种是根据核苷酸序列差异划分的基因型。目前对乙肝基因分型方面的研究表明:HBV的临床特点、预后及治疗结果与HBV的基因型有关,与血清型的关系不明显,所以HBV的基因分型对于临床有重要的指导意义,越来越受到重视。目前临床上对乙肝病毒检测的方法基本是ELISA方法,对乙肝病毒的基因分型通常采用测序的方法,HBV分型试剂正在研究和开发中。 基因芯片的问世为HBV的分型检测提供了一个较好的解决方法,本课题目的在于建立用于HBV病毒检测和分型基因芯片的探针设计和芯片制备方法,建立一个比较完整的基因芯片点样、封闭、杂交和检测的操作体系,并对芯片的特异性、灵敏度和重复性进行测试和分析,对HBV芯片在临床上的应用进行了探索。 实验材料 芯片引物和探针,由上海生工公司合成。 PCR试剂盒,购于华美公司。 临床血清标本来自中国医科大学第一和第二临床附属医院。 仪器:芯片扫描仪(Genomic Solution,Inc),生物芯片点样仪MicroGridⅡ(BioRobotics Lid.),PCR扩增仪(Biometra Personal PCR system,Germany),稳压稳流电泳仪(BIO-RAD MiniⅡ,USA),低温离心机(SIGMA,D-37520,Germany),数显电热恒温干燥箱(202-0,上海阳光实验仪器有限公司)
[Abstract]:The hepatitis B virus (HBV) genome is divided into several subtypes, one of which is the serotype according to the amino acid difference of S region. One is a genotype that is divided according to nucleotide sequence differences. The current studies on HBV genotyping show that the clinical characteristics, prognosis and treatment results of HBV are related to the genotype of HBV, but not to serotype, so the genotyping of HBV has important guiding significance for clinical practice. More and more attention is being paid to it. At present, the detection method of hepatitis B virus in clinic is basically ELISA method. The method of sequencing is usually used for genotyping of hepatitis B virus. HBV typing reagent is being studied and developed. The advent of gene chip provides a good solution for HBV typing detection. The purpose of this project is to establish probe design and chip preparation method for HBV virus detection and typing gene chip. A complete operation system of spot, close, hybridization and detection of gene chip was established, and the specificity, sensitivity and repeatability of the chip were tested and analyzed. The clinical application of HBV chip was explored. The primers and probes were synthesized by Shanghai Bioengineering Company. PCR kit, purchased from Huamei Company. The clinical serum samples were collected from the first and second affiliated hospitals of China Medical University. Instruments: chip scanner (Genomic Solution,Inc), biochip sampler MicroGrid 鈪，
本文编号：2388644
[Abstract]:The hepatitis B virus (HBV) genome is divided into several subtypes, one of which is the serotype according to the amino acid difference of S region. One is a genotype that is divided according to nucleotide sequence differences. The current studies on HBV genotyping show that the clinical characteristics, prognosis and treatment results of HBV are related to the genotype of HBV, but not to serotype, so the genotyping of HBV has important guiding significance for clinical practice. More and more attention is being paid to it. At present, the detection method of hepatitis B virus in clinic is basically ELISA method. The method of sequencing is usually used for genotyping of hepatitis B virus. HBV typing reagent is being studied and developed. The advent of gene chip provides a good solution for HBV typing detection. The purpose of this project is to establish probe design and chip preparation method for HBV virus detection and typing gene chip. A complete operation system of spot, close, hybridization and detection of gene chip was established, and the specificity, sensitivity and repeatability of the chip were tested and analyzed. The clinical application of HBV chip was explored. The primers and probes were synthesized by Shanghai Bioengineering Company. PCR kit, purchased from Huamei Company. The clinical serum samples were collected from the first and second affiliated hospitals of China Medical University. Instruments: chip scanner (Genomic Solution,Inc), biochip sampler MicroGrid 鈪，
本文编号：2388644
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2388644.html
最近更新
教材专著
