Vosberg H P
Max-Planck-Institut für medizinische Forschung, Heidelberg, Federal Republic of Germany.
Hum Genet. 1989 Aug;83(1):1-15. doi: 10.1007/BF00274139.
The polymerase chain reaction (PCR) is a method for the selective amplification of DNA or RNA segments of up to 2 kilobase-pairs (kb) or more in length. Synthetic oligonucleotides flanking sequences of interest are used in repeated cycles of enzymatic primer extension in opposite and overlapping directions. The essential steps in each cycle are thermal denaturation of double-stranded target molecules, primer annealing to both strands and enzymatic synthesis of DNA. The use of the heat-stable DNA polymerase from the archebacterium Thermus aquaticus (Taq polymerase) makes the reaction amenable to automation. Since both strands of a given DNA segment are used as templates, the number of target sequences increases exponentially. The reaction is simple, fast and extremely sensitive. The DNA or RNA content of a single cell is sufficient to detect a specific sequence. This method greatly facilitates the diagnosis of mutations or sequence polymorphisms of various types in human genetics, and the detection of pathogenic components and conditions in the context of clinical research and diagnostics; it is also useful in simplifying complex analytical or synthetic protocols in basic molecular biology. This article describes the principles of the reaction and discusses the applications in different areas of biomedical research.
聚合酶链反应（PCR）是一种用于选择性扩增长度达2千碱基对（kb）或更长的DNA或RNA片段的方法。在相反且重叠的方向上，利用与目标序列侧翼的合成寡核苷酸进行酶促引物延伸的重复循环。每个循环的基本步骤是双链目标分子的热变性、引物与两条链的退火以及DNA的酶促合成。使用来自嗜热栖热菌的热稳定DNA聚合酶（Taq聚合酶）使该反应适合自动化操作。由于给定DNA片段的两条链都用作模板，目标序列的数量呈指数增加。该反应简单、快速且极其灵敏。单个细胞的DNA或RNA含量就足以检测特定序列。这种方法极大地促进了人类遗传学中各种类型突变或序列多态性的诊断，以及临床研究和诊断中致病成分和病症的检测；它在简化基础分子生物学中的复杂分析或合成方案方面也很有用。本文描述了该反应的原理，并讨论了其在生物医学研究不同领域的应用。
