HBV基因分型芯片的构建及临床初步应用

发布时间：2018-01-05 08:50

本文关键词：HBV基因分型芯片的构建及临床初步应用　出处：《中国医科大学》2006年硕士论文　论文类型：学位论文

【摘要】：背景及目的乙肝病毒(Hepatitis B Virus,HBV)感染呈世界性分布,其分型有血清型和基因型两种。它是一种高变异病毒,目前根据其基因组核苷酸序列差异已将HBV基因型分为A-H型。由于传统的血清学分析不能真正反映基因组的改变,不能反映HBV的进化关系及变异情况,而基因型在近年的研究中发现其与病毒的热点突变、疾病的活动度、抗病毒药物的疗效及疾病的预后等都有一定相关性,所以鉴别基因分型对流行病学调查与指导个性化治疗具有十分重要的意义,因此开发简便、有效的基因分型方法有很大的实用价值。基因芯片的问世为HBV分型检测提供了一个较好的解决办法,其高通量、并行化检测的特点在HBV分型方面具有独特的优势。目前国内对基因芯片的报道诸多,但尚没有成熟的HBV芯片应用到临床,并且我国的基因型研究开展较少,所以本研究使用本实验室建立的技术流程自行构建HBV基因分型芯片,着重解决分型探针的设计问题,优化杂交条件,对芯片的重复性与可靠性进行检测分析,并应用于临床,对150例不同类型的乙肝患者进行HBV基因分型,为本地HBV基因型分布提供调查资料。实验材料与方法一、实验材料临床血清标本来自中国医科大学第二临床医院传染科住院患者。 PCR试剂盒(华美生物工程公司) 引物与探针(上海博亚生物工程公司合成) PCR扩增仪(Biometra Personal PCR system,Germany)生物芯片点样仪(Micro Grind Ⅱ600,Biorobtics Ltd,England)
[Abstract]:Background and purpose
Hepatitis B virus (Hepatitis B, Virus, HBV) infection is a worldwide distribution, the type of serotype and genotype in two. It is a high variability of the virus, according to the nucleotide sequence of the HBV gene type differences have been divided into A-H type. Because the traditional serological analysis can not really reflect the change of genome that does not reflect the evolution and variation of HBV, and the genotype in recent studies found that mutations in the virus and hot, disease activity, disease curative effect and prognosis of antiviral drugs have some relevance, so the identification of genotypes is very important for epidemiological investigation and guiding personalized therapy therefore the development of simple and effective genotyping method has great practical value.
The advent of gene chip provides a good solution for HBV genotyping, the high flux, has a unique advantage of parallel detection in HBV typing. At present, many reports of gene chips, but there is no mature HBV chip applied to the clinical and research of genotype in China carry out less, so the laboratory technology process establishment constructed HBV genotyping chip used in this study, focus on solving the design problem type probe, optimization of hybridization conditions, the repeatability and reliability of the chip were analyzed, and should be used clinically in 150 cases of hepatitis B patients with different HBV genotypes with the survey data, the local distribution of HBV genotypes.
Experimental materials and methods
First, experimental materials
The clinical serum specimens are from the inpatients in the infectious department of the Second Clinical Hospital of China Medical University.
PCR Kit (Huamei bioengineering company)
Primers and probes (Shanghai Boya bio engineering company synthesis)
PCR amplification instrument (Biometra Personal PCR system, Germany) biochip dot sampler (Micro Grind II 600, Biorobtics Ltd, England)

【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346

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