传染病病原诊断技术.ppt

A Novel Coronavirus Associated with Severe Acute Respiratory Syndrome virus-isolation techniques electron-microscopical histologic studies Molecular and serologic assays Primer pair IN-2 (+) 5GGGTTGGGACTATCCTAAGTGTGA3 IN-4 (–) 5TAACACACAACICCATCATCA3 conserved regions of open reading frame 1b A 405-nucleotide segment The Genome sequence of the SARS-associated coronavirus Science 300 (5624), 1399-1404 (2003) Virus isolation ( bronchoalveolar lavage specimen) Viral particles were purified, genetic material (RNA) was extracted RNA was converted to cDNA by random-priming and oligo(dT) primings trategy Size-selected cDNA products were cloned single sequence reads Sequences were assembled Rapid amplification of cDNA ends [RACE] to capture the 5 end of the viral genome. 病毒性感染样本（咽拭子，痰，肺泡灌洗液，脑脊液） 保存活病毒样本 核酸抽提、逆转录 多重PCR扩增和质谱分析 阳性 阴性 部分样品进行病毒培养 阴性 阳性 部分样品进行基因芯片检测 阴性 阳性 病毒扩增，电子显微镜观察，病毒核酸的分子克隆，序列的比对，进化分析以及基因功能的初步探讨。 DNaseI处理----Random PCR 克隆扩增DNA，大规模测序，核酸序列比对，进化树分析。 不明原因感染的病原体筛查系统 * * * 用ELISA和liquichip 对硫氧还蛋白进行定量测定得到的标准曲线　　I: 使用liquichip测定得到的标准曲线　　II、III：使用ELISA测定得到的标准曲线　　可以看出liquichip的灵敏度更高，在更低的蛋白质浓度下，曲线即呈现线性趋势。 * さらにアジレントでは最初にお話しましたように、アレイ以外でも遺伝子発現解析に必要な全てのツールを提供しています。個々の製品の詳しい説明については、ぜひアジレントのブースまでお越しください。 カタログアレイ→Cdnaカタログアレイに変える * Agilent not only has the manufacturing flexibility to produce different types of microarrays, but also has developed a Web-based custom array design tool to enable customers to take advantage of the flexibility of our platform. eArray currently offers various array layouts. Customers can design custom arrays built from probes and probe sets used in our catalogue arrays, or sources of their own, or a mix and match of both. We have added a new functionality on eArray to allow scientists to collaborate and work on a microarrays design as a group. This has added benefits to our customers. In the next coming months, eArray will include other advanced features and functionality to make it a even more powerful tool for custom ar