Applications of PCR detection
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What is PCR testing?
Polymerase chain reaction (PCR) uses DNA fragments as templates, with the participation of DNA polymerase and nucleotide substrates, to amplify DNA to a sufficient amount for structural and functional analysis. PCR detection method is of great significance for the rapid diagnosis of clinical bacterial infectious diseases.
What is the principle of PCR?
The principle of PCR is to amplify DNA fragments located between two known sequences, similar to the natural DNA replication process. Taking the DNA molecule to be amplified as a template, and a pair of oligonucleotide fragments complementary to the 5' end and 3' end of the template as primers, under the action of DNA polymerase, according to the semi-reserved replication mechanism of the template, the chain is separated. Extend until new DNA synthesis is complete and repeat the process to amplify the target DNA fragment.
Applications of PCR detection:
1. Pathogen detection
Currently, fluorescent quantitative PCR detection technology can detect gonococcus, Chlamydia trachomatis, Ureaplasma urealyticum, human papilloma virus, herpes simplex virus, human immunodeficiency virus, hepatitis virus, influenza virus, Mycobacterium tuberculosis, eb virus and cytomegalo Quantitative determination of pathogens such as viruses. Compared with traditional detection methods, it has the advantages of high sensitivity, less sampling, fast and convenient.
2. Prenatal diagnosis
So far, there is no treatment for genetic diseases caused by changes in genetic material. Only prenatal monitoring can reduce the birth of sick babies and prevent the occurrence of various genetic diseases. For example, reducing the number of children with an X-linked genetic disorder is a non-invasive approach that is easily accepted by pregnant women.
3. Evaluation of drug efficacy
Quantitative analysis of hepatitis B virus (HBV) and hepatitis C virus (HCV) has shown that the amount of virus correlates with the efficacy of certain drugs. High levels of HCV expression are insensitive to the effects of interferon, whereas low titers of HCV are sensitive to the effects of interferon; serum levels of HBV-DNA have decreased briefly during lamivudine treatment, and if increased again or more than before level, indicating that the virus has mutated. Such as the application and significance of PCR technology in HBV detection:
(1) Know the amount of hepatitis B virus in the body.
(2) Whether to copy.
(3) Whether it is contagious, and how contagious it is.
(4) Whether it is necessary to take medicine.
(5) Whether abnormal liver function is caused by virus.
(6) Determine which antiviral drug is suitable for the patient.
(7) Determine the efficacy of drug treatment.
4. Tumor gene detection
Although tumor pathogenesis remains unclear, it is widely accepted that mutations in related genes are potential causes of oncogenic transformation. In many tumors, increased expression and mutations of oncogenes can appear at an early stage. Real-time quantitative PCR can not only effectively detect gene mutations, but also accurately detect the expression of oncogenes. At present, this method has been used to detect the expression of telomerase hTERT gene, chronic myeloid leukemia WT1 gene, tumor ER gene, prostate cancer PSM gene, tumor-associated virus gene and other genes. With the continuous discovery of new tumor-related genes, real-time PCR technology will play a greater role in tumor research.
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