What is PCR?
Quantitative Real-time PCR (Quantitative Real-time PCR) is a method for measuring the total amount of product after each polymerase chain reaction (PCR) cycle with fluorescent chemicals in a DNA amplification reaction. A method for quantitative analysis of specific DNA sequences in a sample to be tested by an internal or external reference method. ·
Real-time PCR is a real-time detection of the PCR process through fluorescent signals during the PCR amplification process. Since there is a linear relationship between the Ct value of the template and the initial copy number of the template in the exponential period of PCR amplification, it becomes the basis for quantification.
The principle of PCR technology
After the Taqman probe labeled with fluorescein is mixed with the template DNA, the thermal cycle of high temperature denaturation, low temperature renaturation, and suitable temperature extension is completed, and the Taqman probe that is complementary to the template DNA is cut off according to the law of polymerase chain reaction. Fluorescein is free in the reaction system and emits fluorescence under specific light excitation. With the increase of the number of cycles, the amplified target gene fragments increase exponentially. By real-time detection of the corresponding fluorescence signal intensity that changes with the amplification , obtain the Ct value, and use several standards with known template concentrations as controls to obtain the copy number of the target gene in the sample to be tested.
Application of PCR technology in clinical disease diagnosis
Diagnosis and efficacy evaluation of various infectious diseases such as hepatitis, AIDS, avian influenza, tuberculosis, and sexually transmitted diseases; eugenics and prenatal testing for thalassemia, hemophilia, gender dysplasia, mental retardation syndrome, and fetal malformation; tumor marker and tumor gene detection Realize the diagnosis of tumor diseases; genetic testing realizes the diagnosis of genetic diseases.
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