Primer group for multiple RPA detection of respiratory tract pathogens, detection reagent and kit

Abstract

The invention provides a primer group for multiple RPA detection of respiratory tract pathogens, a detection reagent and a kit. The high-specificity primer group for multiple RPA detection is successfully obtained through RPA primer design improvement of at least 5 respiratory tract pathogens of human cytomegalovirus, adenovirus, parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3,respiratory syncytial virus, staphylococcus aureus, streptococcus pneumoniae, mycoplasma pneumoniae, chlamydia pneumoniae, klebsiella pneumoniae, acinetobacter baumannii, pseudomonas aeruginosa, stenotrophomonas maltophilia and legionella pneumophila, the detection reagent is prepared with the adoption of the primer group, and the kit is prepared, so that RPA detection of up to 15 respiratory tract pathogens can be realized according to speculation, time required for the whole detection procedure can be shortened, and steps are simplified.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a primer group, a detection reagent and a kit for multiple RPA detection of respiratory pathogens. Background technique [0002] RPA technology (full name "recombinase-mediated amplification technology") is known as a nucleic acid detection technology that can replace PCR technology. RPA technology is a technology that can rapidly amplify nucleic acid at a constant temperature. RPA technology mainly relies on three enzymes: recombinase that can bind primers, single-stranded DNA binding protein (SSB) and DNA polymerase; the optimal reaction temperature of the mixture of these three enzymes is around 37°C. The principle of RPA technology is: the protein-DNA complex formed by the combination of recombinase and primer can search for homologous sequences in double-stranded DNA. With the help of the double-stranded DNA template, the primers are paired with the homologous sequence, an...

AI Technical Summary

Problems solved by technology

Since RPA technology is usually carried out under isothermal conditions, it cannot avoid the differences in Tm values ​​and amplification efficiencies between different respiratory pathogen primer sets during multiple detection by adjusting the annealing temperature and cooling rate during the reaction like ordinary PCR amplification. The mutual interference between various amplification products brought about also makes it very difficult to obtain highly specific RPA amplification primers suitable for multiple detection

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