HBV基因芯片的制備及其在臨床應(yīng)用的研究
發(fā)布時(shí)間：2018-12-21 08:14
【摘要】：前言 乙肝病毒(HBV)的感染呈世界性的分布,乙肝病毒基因組分為多個(gè)亞型,一種是根據(jù)S區(qū)的氨基酸差異劃分的血清型,一種是根據(jù)核苷酸序列差異劃分的基因型。目前對(duì)乙肝基因分型方面的研究表明:HBV的臨床特點(diǎn)、預(yù)后及治療結(jié)果與HBV的基因型有關(guān),與血清型的關(guān)系不明顯,所以HBV的基因分型對(duì)于臨床有重要的指導(dǎo)意義,越來越受到重視。目前臨床上對(duì)乙肝病毒檢測(cè)的方法基本是ELISA方法,對(duì)乙肝病毒的基因分型通常采用測(cè)序的方法,HBV分型試劑正在研究和開發(fā)中。 基因芯片的問世為HBV的分型檢測(cè)提供了一個(gè)較好的解決方法,本課題目的在于建立用于HBV病毒檢測(cè)和分型基因芯片的探針設(shè)計(jì)和芯片制備方法,建立一個(gè)比較完整的基因芯片點(diǎn)樣、封閉、雜交和檢測(cè)的操作體系,并對(duì)芯片的特異性、靈敏度和重復(fù)性進(jìn)行測(cè)試和分析,對(duì)HBV芯片在臨床上的應(yīng)用進(jìn)行了探索。 實(shí)驗(yàn)材料 芯片引物和探針,由上海生工公司合成。 PCR試劑盒,購(gòu)于華美公司。 臨床血清標(biāo)本來自中國(guó)醫(yī)科大學(xué)第一和第二臨床附屬醫(yī)院。 儀器:芯片掃描儀(Genomic Solution,Inc),生物芯片點(diǎn)樣儀MicroGridⅡ(BioRobotics Lid.),PCR擴(kuò)增儀(Biometra Personal PCR system,Germany),穩(wěn)壓穩(wěn)流電泳儀(BIO-RAD MiniⅡ,USA),低溫離心機(jī)(SIGMA,D-37520,Germany),數(shù)顯電熱恒溫干燥箱(202-0,上海陽光實(shí)驗(yàn)儀器有限公司)
[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，
本文編號(hào)：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，
本文編號(hào)：2388644
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