Real Time PCR研究进展及其在海洋病原生物检测中的应用

Development of Real Time PCR and application in detection of marine pathogenic organism

Real Time PCR,即实时监测PCR扩增产物并进行解析的方法,目前已广泛应用于分子生物学研究的各个领域。Real Time PCR技术秉承及发展了普通PCR的快速、高灵敏度检出等优点,同时克服了普通PCR不能准确定量、容易污染等缺点,无需在反应结束后通过电泳操作确认扩增产物。目前,Real Time PCR可设计多对引物在同一反应体系中同时对多个靶基因进行扩增,实现多重实时定量检测。Real Time PCR使PCR技术发生了质的飞跃,扩展了PCR技术的应用范畴,是一种具有划时代意义的技术。本文主要介绍Real Time PCR的主要原理、解析方法、技术发展趋势及其在海洋病原生物检测方面的应用。

The Real Time PCR can be used to monitor timely and increase the information of DNA, therefore it is applied in the fields. The Real-Time chemistries allow for the detection of PCR amplification during the early phases of the reaction. The measure of the ki- netics of the reaction in the early phases of PCR provides a distinct advantage over traditional PCR detection. The Real-Time PCR de- tects the accumulation of amplification during the reaction. The data are then measured at the exponential phase of the PCR reaction. It has become one of the most widely used methods to assess the gene expression owing to its sensitivity and reduce the run time compared with the traditional methods. Several different types of real-time PCR are being marketed to the scientific community at this time, each with their advantages. In the present paper, the principle and application in detection of marine pathogenic organism of real time PCRtechnique are described.