How does the fluorescent PCR method perform nucleic acid detection?

PCR laboratory is also called gene amplification laboratory. Real-time fluorescent quantitative PCR technology is a leap forward in DNA quantitative technology. It is used to amplify specific DNA fragments, which can be regarded as special DNA replication in vitro. Through the DNA gene tracking system, the virus content in the patient's body can be quickly grasped with an accuracy of nanometer level. The fluorescent PCR method is the carrier that realizes this technology. Through the method of amplifying the genes contained in the virus, it can detect whether some infected persons with low virus content contain specific viruses.

The PCR laboratory is actually extremely powerful. Qualitative analysis and quantitative detection are its two biggest application directions. The application in the detection of new crown nucleic acid is particularly prominent. Next, Desheng will introduce to you the application of PCR in the detection of pathogens. I hope that through these examples, you can better understand what the nucleic acid detection process is.

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Example: Novel coronavirus nucleic acid detection.

Methodology: fluorescence PCR method.

Uses: Used for in vitro qualitative detection of suspected cases of pneumonia, suspected clusters of patients with new coronavirus infection, and other patients who need to diagnose or identify new coronavirus infection.

Detection gene: new coronavirus (2019-nCoV) ORFlab and N gene, E gene.

Specimen types: upper respiratory tract specimens: throat swabs, nasal swabs; lower respiratory tract specimens: respiratory tract extracts, alveolar lavage fluid samples, lung tissue biopsy specimens.

Detection process:

(1) Biosafety protection before nucleic acid detection (personal three-level protective wear by staff) → (2) Inactivation of samples (inactivation at 56°C for 30 minutes) → (3) Nucleic acid extraction of samples (manual, nucleic acid extractor) )→(4) PCR system preparation and template loading→(5) On-board testing and result analysis→(6) Disinfection after testing.

Other pathogen detection: HIV, hepatitis A, B, C, HPV, lung branches, lung coat, Epstein-Barr virus, adenovirus, cytovirus, A and B stream, Ebola virus, group B streptococcus.

Hospitals and departments suitable for development: children's hospitals, maternity and children's clinics, pediatric clinics, third parties, hospitals of various levels in need

The advent of PCR technology enables rapid and convenient pathogen detection. The PCR detection method has the advantages of high sensitivity, high specificity, quickness, and low sample requirements. Therefore, it is widely recognized by clinicians and has been widely used in clinical diagnosis in hospitals and the diagnosis of poultry disease in various epidemic prevention and detection departments.