Fluorescent RT-PCR reagent and method for detecting influenza A virus, influenza B virus and coronavirus SARS-CoV-2

Abstract

The invention provides a freeze-dried type multiple fluorescent RT-PCR reagent and a method for detecting and distinguishing influenza A virus, influenza B virus and coronavirus SARS-CoV-2. Primers and probes of the fluorescent RT-PCR reagent for detecting the influenza A virus, the influenza B virus and the coronavirus SARS-CoV-2 provided by the invention are obtained by designing and screening based on the latest gene sequence information of the influenza A virus, the influenza B virus and the coronavirus SARS-CoV-2 in a public gene sequence database, and the reagent has good inclusivity, specificity and reaction performance. According to the reagent, a human RNase P nucleic acid fragment is used as a detection internal reference. In addition, the fluorescent RT-PCR detection reagent forthe influenza A virus, the influenza B virus and the coronavirus SARS-CoV-2 is prepared into a freeze-dried form capable of being stored at normal temperature through freeze-drying technology, thus avoiding the risk that the conventional liquid fluorescent PCR detection reagent is invalid due to the change of storage temperature or repeated freezing and thawing, and the transportation and management cost is reduced.

Description

technical field [0001] The invention relates to pathogenic microorganism detection technology, in particular to nucleic acid detection of influenza A virus, influenza B virus and coronavirus SARS-CoV-2. Background technique [0002] Respiratory tract infection is the most common type of disease in humans, and it is one of the main causes of morbidity and death in the population. Research shows that respiratory viruses are the most common pathogens causing respiratory diseases. Respiratory viruses include influenza virus, coronavirus, rhinovirus, respiratory syncytial virus, parainfluenza virus, human metapneumovirus, adenovirus, etc. The clinical manifestations caused by respiratory virus infection are relatively similar, mainly rhinitis, pharyngitis, laryngitis, tonsillitis, tracheitis, bronchitis, pneumonia, cough, dyspnea, fever, fatigue, etc., but the infection caused by different pathogens, the treatment Methods, efficacy and course of disease are not the same. [00...

AI Technical Summary

Problems solved by technology

However, conventional fluorescent RT-PCR detection reagents are all liquid reagents, which need to be stored below -20°C to maintain detection activity. Temperature rise and repeated freezing and thawing during storage, transportation and use will reduce the detection performance of the reagents

Especially during the epidemic period, transportation is blocked, cold chain transportation conditions are difficult to guarantee, the risk of reagent failure and cold chain management costs greatly increase during long-distance transportation, and it cannot adapt to the application of international long-distance transportation and grass-roots testing institutions with poor cold chain conditions.

[0006] In addition, due to the outbreak of the global epidemic, different regions entered the winter influenza season separately, which brought greater challenges to epidemic control due to the similarity of early symptoms between influenza and COVID-19

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