MIA primer, probe and kit for detecting COVID-19 and application of MIA primer and probe

Abstract

The invention discloses an MIA primer, a probe and a kit for detecting COVID-19 and an application of the MIA primer and the probe. According to the invention, the MIA primer and the probe are respectively designed based on a nucleocapsid N gene, an envelope E gene and an ORF1ab region of the COVID-19 publicized by NCBI. According to the MIA method disclosed by the invention, the detection of ORF1ab, E and N target genes of COVID-19 is realized in 3-15 minutes for the first time, and the detection sensitivity reaches 100 copies / ml (sample template concentration). The MIA method has no non-specific cross reaction with pathogen nucleic acid samples such as type A influenza, type B influenza and respiratory syncytial virus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to MIA primers, probes, kits and applications for detecting novel coronavirus pneumonia. Background technique [0002] Corona Virus Disease 2019 (COVID-19), referred to as "New Coronary Pneumonia", named by the World Health Organization as "2019 Coronavirus Disease", refers to the pneumonia caused by 2019 Novel Coronavirus infection. The clinical manifestations of pneumonia patients infected by the new coronavirus are: fever, fatigue, dry cough as the main manifestations, nasal congestion, runny nose and other upper respiratory symptoms are rare, and hypoxic and hypoxic states will occur. About half of the patients developed dyspnea more than a week later, and severe cases could rapidly progress to acute respiratory distress syndrome, septic shock, difficult-to-correct metabolic acidosis, and coagulation dysfunction. According to the "New Coronavirus Pneumonia Diagnosis and...

AI Technical Summary

Problems solved by technology

According to the "New Coronavirus Pneumonia Diagnosis and Treatment Plan (Trial Seventh Edition)" document, nucleic acid testing is still the preferred option for diagnosis. In this document, for the first time, positive serum novel coronavirus-specific IgM antibodies and IgG antibodies were added as a confirmed diagnosis. However, since the antibodies produced by the human body are in the middle and late stage of disease development, early invisible carriers cannot be diagnosed and screened by antibody detection, so this method can generally only be used as an auxiliary detection method other than nucleic acid detection.

Clinical CT images and gene sequencing are also an important basis for doctors to judge whether a patient is infected, but due to the scarcity of equipment and the complexity of diagnosis, it is difficult to popularize

[0003] Fluorescent quantitative PCR is the preferred solution for the detection of the new coronavirus at this stage, but it has the following limitations: (1) It is easy to cause false positives due to aerosol pollution, and professionally trained technicians with PCR certificates, professional PCR certification laboratories and Expensive analysis equipment; (2) Traditional fluorescence quantitative PCR sample nucleic acid extraction and amplification detection are carried out in two steps, and it takes more than 4 hours from sampling to detection results

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