Real-time quantitative polymerase chain reaction (Real Time PCR, if used for RNA detection, which is called reverse transcription real-time PCR, Real-time RT-PCR) is a real-time PCR method, which refers to DNA. Or reverse-input (RT-PCR) RNA is detected by polymerase chain reaction and real-time monitoring of DNA amplification process, and amplification products are measured during exponential growth of amplification because of amplification of exponential growth phase measurements and specific DNA There is a correlation between the (RNA) starting amounts to achieve quantitative detection.
The basic goal of Real Time PCR is to accurately measure and identify very small amounts of specific nucleic acids, so that the amount of the original target gene can be quantified by monitoring the CT value. A variety of conventional PCR methods currently in common use, such as various gel electrophoresis PCR methods, end-point fluorescence quantification or PCR-ELISA are called end-point PCR methods. The biggest advantage of real-time PCR is to overcome the large error of quantification of the end-point PCR method after entering the plateau or saturation period, and to achieve accurate quantification of DNA/RNA.
Ordinary PCR technology was invented in 1983, and real-time fluorescent quantitative PCR technology was invented in the 1990s and produced the world's first real-time fluorescence quantitative PCR instrument in 1996. The technical elements realized by real-time fluorescent quantitative PCR technology are mainly Fluorescence quantitative PCR kit and real-time fluorescent quantitative PCR instrument, China's fluorescent quantitative PCR kit has strong research and development strength, and there are many manufacturers. The clinically diagnosed varieties such as HBV/HCV and avian influenza diagnostic kit have excellent performance and sufficient supply. Localization. In the case where the swine influenza virus RNA nucleic acid sequence is clear, the human infection swine influenza real-time fluorescent quantitative PCR diagnostic kit will soon be available using the existing reagent development platform. Due to the high technical content of real-time PCR instruments, only a few Chinese companies have the ability to produce TL-988 series instruments such as Xi'an Tianlong Technology Co., Ltd., which not only obtained invention patents, national science and technology awards, national key new product certificates, The medical device registration certificate has been established by the National Development and Reform Commission as the national high-tech industrialization demonstration project in 2008. The product performance has reached the international advanced level, and the market share in the dairy testing industry is higher than that of imported instruments [1-3]. Real-time PCR machines are widely used. Since 1999, Western developed countries have tried to apply PCR to clinical infectious disease diagnosis, food safety testing and blood screening. On December 13, 2008, the Ministry of Health of China issued the "Health Emergency Team Equipment Reference Catalogue" (Wei Ban Emergency Development [2008] No. 207), which requires the establishment of a PCR instrument and a real-time fluorescence quantitative PCR instrument in the construction of a health emergency team above the county level. As an infectious disease control equipment.
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