关键词: 实时荧光定量PCR, 原理, 应用, 进展, 综述文献

Abstract:

BACKGROUND: Real-time fluorescence quantitative polymerase chain reaction (PCR) refers to join the fluorescence groups into PCR reacting system, and to real-time monitor entire PCR process using the fluorescence signal accumulation, finally to make the quantitative analysis of the unknown template through the standard curve.
OBJECTIVE: To study the theory of real-time fluorescence quantitative PCR and to explore its applications and progress in medicine.
METHODS: With “real-time fluorescence quota PCR, theorem, application” in English for the search term, PubMed database was retrieved from January 2000 to December 2008. With “real-time fluorescence quantitative PCR, principle, application” in Chinese for the search term, Wanfang Database from January 2000 to December 2008, Tsinghua Tongfang Chinese database from January 2000 to December 2008 were was retrieved. Literatures were limited to English and Chinese languages. Cell factor and tumor resistance genes served as the evaluation index. The methodology of research on the real-time fluorescence quantitative PCR technology and medical applied research on real-time fluorescence quantitative PCR technology were included. While repetitive research and Meta analysis were excluded.
RESULTS AND CONCLUSION: Because real-time fluorescence quantitative PCR technology has not only realized PCR develops from qualitative to quantitative levels, but also has strong specificity, high sensitivity, good duplication, accurate quantization, high automaticity, and entire blocking response compared with conventional PCR, thus it becomes the important tool in the molecular biology research. Real-time fluorescence quantitative PCR technique has been widely applied, such as mRNA expression, detections of DNA copy number and determination of mononucleotide polymorphism, as well as in the clinical medicine including accurate quantitative examination of mycobacterium tuberculosis, Type B and Type C hepatitis, AIDS virus, gonococcus, and chlamydia trachomatis. Its quantitative scope extremely extends, no need of gradient dilution, the specificity is stronger, overcomes the false positivity. Due to the traditional PCR technology cannot give the accurate quantization, it is greatly limited in the practical application. Therefore, the accurate quantization of the PCR product, particularly the dynamic monitoring of viral etiology, becomes the urgent need.