What is PCR testing?
Introduction of PCR
Polymerase chain reaction ( PCR) uses a piece of DNA as a template to amplify the DNA to a sufficient amount for structural and functional analysis with the participation of DNA polymerase and a nucleotide substrate. PCR detection method is of great significance in the clinical rapid diagnosis of bacterial infectious diseases.
The principle of PCR is used to amplify DNA fragments located between two known sequences, similar to the replication process of natural DNA. Using 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 mechanism of semi-reserved replication along the template The chain is extended until new DNA synthesis is completed, and this process is repeated to amplify the target DNA fragment.
The usage of PCR testing:
1. Pathogen detection
At present, 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, Epstein-Barr virus and cytomegalo Quantitative determination of pathogens such as viruses. Compared with the traditional detection method, it has the advantages of high sensitivity, less sampling, quick and easy and so on.
2. Prenatal diagnosis
So far, there is no cure for hereditary diseases caused by changes in hereditary material. Only prenatal monitoring can be used to reduce the birth of sick babies to prevent the occurrence of various hereditary diseases. For example, to reduce the number of children with X-linked genetic diseases It is a non-invasive method that is easily accepted by pregnant women.
3. Drug efficacy assessment
Quantitative analysis of hepatitis B virus (HBV) and hepatitis C virus (HCV) showed that the amount of virus was related to the curative effect of some drugs. The high level of HCV expression is not sensitive to the effect of interferon treatment, while the low titer of HCV is sensitive to the effect of interferon; during the treatment of lamivudine, the serum content of HBV-DNA once decreased, and then increased again or exceeded the previous level if , 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 changes in liver function are caused by viruses.
(6). Determine which type of antiviral drug is suitable for the patient.
(7). Determine the efficacy of drug treatment.
4. Tumor gene detection
Although the mechanism of tumor pathogenesis remains unclear, it is widely accepted that mutations in related genes are the underlying cause of oncogenic transformation. Increased expression and mutation of oncogenes can appear early in many tumors. Real-time fluorescence 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 various genes such as telomerase hTERT gene, chronic myeloid leukemia WT1 gene, tumor ER gene, prostate cancer PSM gene, tumor-related virus gene and so on. With the continuous discovery of new genes related to tumors, real-time PCR technology will play a greater role in tumor research.
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