《生命科学》 2020, 32(5): 515-521
焦磷酸光化测序技术的发展和应用
摘 要：
摘 要：人类基因组计划(Human Genome Project， HGP) 不仅极大地提高了人类对基因组和相关遗传信息的认识水平，而且促进了生命科学研究技术的发展和应用。正是在这样的历史背景下，焦磷酸光化测序技术(pyrosequencing) 由瑞典研究人员发明。焦磷酸光化测序技术的基础原理是基于“通过合成测序”原理进行酶促反应的DNA 测序方法，通过基于释放焦磷酸盐时的链式反应的可见光检测，即可获得一个特异的检测峰，峰值的高低和相匹配的碱基数成正比。该技术可应用于DNA 核苷酸序列和突变的检测、单核苷酸多态性的基因型的鉴定，以及DNA 甲基化水平变化的分析等。近年来，随着摄影器材和成像技术的快速发展，这项技术的原理是基于通过酶促反应而实时检测可见光，因此，有望在检测的敏感性方面得到更进一步的发展。该文根据笔者在瑞典近三十年的工作经验和积累的文献，首先阐明焦磷酸光化测序的基本原理，然后介绍该技术的应用，最后讨论其发展前景。
Abstract:
Abstract: The Human Genome Project (HGP) not only greatly improved the understanding of human genome and related genetic information， but also promoted the development of technologies in life science research. It was under this historical context that pyrosequencing was invented by Swedish researchers. Pyrosequencing is a method of DNA sequencing based on the "sequencing by synthesis" principle with enzymatic reactions， and relies on light detection based upon a chain reaction when pyrophosphate is released. The application of this technology involved the detection of DNA nucleotide sequences and mutations， the identification of genotypes of single nucleotide polymorphisms， the analysis of changes in DNA methylation levels etc. With the recent rapid development of photographic equipment and imaging technology， this technology is expected to have an increasing sensitivity in signal detection. Based upon the working experiences in Sweden for nearly 30 years and accumulated literature， this review first clarified the basic principles of pyrosequencing， then introduced the applications of this technology， and finally discussed its development prospects in the near future.
