病原体分子诊断技术进展.pptVIP

Nucleic Acids can be single- or double-stranded. Double stranded DNA or dsDNA is shown in the top picture. The 2 strands are held together by hydrogen bonds between complementary bases on the strands. Complementary bases of 2 nucleotides are called base pairs: C pairs with G A pairs with T (which is in DNA only) A pairs with U (which is in RNA only) ? Typically, DNA is found in the double stranded DNA form, known as the characteristic DNA double helix, but can also exist in single stranded form. RNA commonly exists as a single-stranded molecule. In most bacterial cells, there are several thousand copies of ribosomal RNA (rRNA) per organism. This is in contrast to DNA that is usually only present in one or a few copies per cell. This is why Gen-Probe targets the ribosomal RNA, because there is more of it available in each cell. The technology of targeting ribosomal RNA was among the first patented technologies developed by Gen-Probe. (Read from Slide) This is an example scenario. The basic component in Gen-Probe’s Molecular Diagnostics Assays is a nucleic acid probe made of DNA. (Read bullet points from slide) When the probe binds with the target, the detector molecule on the probe can indicate when the probe has hybridized with its target. * 病原体分子诊断技术进展 分子诊断概念和特点 概念：利用标本中的核酸作为检测对象，对感染性疾病、肿瘤、遗传性疾病等进行诊断和预后判断的方法。 特点 特异性高、灵敏度高 稳定性高 诊断范围广、适应性强 临床应用前景好 什么是核酸？ 链状分子 DNA (脱氧核糖核酸) 贮存遗传信息 RNA (核糖核酸) 传递遗传信息 遗传信息的传递途径 DNA RNA 蛋白质 转录 逆转录 DNA 和 RNA 的区别 糖基 + Base Nucleoside + Phosphate P Nucleotide A、DNA和RNA的主要区别就在骨架上的糖基。 B、DNA很稳定，RNA不稳定。 RNA 和 DNA 的结构 dsDNA ssDNA ssRNA 通常RNA在细胞中拷贝数远高于DNA的拷贝数 一个或几个拷贝DNA 可达10000个拷贝的核糖体RNA (rRNA) One Cell Contains: 如使用rRNA为靶标，将极大地提高了取样效率和扩增检测灵敏度 A、如果2ml样本含有1个细菌 取0.2ml样本，最终得到细菌的几率是1/10。 Sample Cell Lysis Target RNA Released B、如果2ml样本含有1个细菌 先裂解，后取样， 2ml样本含有10000个rRNA。 取0.2ML样本，可得到 1000个分子 主要的核酸扩增技术 DNA扩增技术 A、PCR：使用Taq酶进行扩增，温度循环；RT-PCR将RNA变成DNA，再进行DNA扩增 B、