RPA (recombinase polymerase amplification) method, PRA special primer and RPA probe for detecting universal human adenovirus, and application of RPA probe

Abstract

The invention provides an RPA (recombinase polymerase amplification) method, an PRA special primer and an RPA probe for detecting universal human adenovirus, and application of the PRA special primerand RPA probe in detection of the universal human adenovirus. The detection method, the special primer and the probe are designed based on a common conserved sequence of type 3, type 7, type 11, type21 and type 55 human adenovirus hexon genes, and have the oligonucleotide sequences shown as SEQ ID NO. 3, SEQ ID NO. 4 and SEQ ID NO. 5. According to the method, the novel isothermal amplification technology is applied to detection of the universal human adenovirus for the first time; the enzymatic reaction process of in-vivo DNA replication is simulated, a specific enzyme and protein combination(recombinase, single-stranded binding protein and DNA polymerase) is used for DNA template amplification, amplification of a specific nucleotide sequence can be realized at a constant temperature of37 DEG C, and amplification products can be visually judged through a lateral chromatographic test strip. The detection method has high sensitivity, and the detection limit can reach 101 copies / reaction; the specificity is high, and no cross-reaction with other pathogens exists; the detection speed is high, and the method is simple and convenient to operate.

Description

technical field [0001] The invention relates to an RPA method for detecting a general-purpose human adenovirus, its special primers and probes, and uses thereof, which belong to the field of biotechnology, relate to the molecular biology of a general-purpose human adenovirus (human adenovirus, HAdV), and relate to a detection The method and its use of universal human adenovirus, especially a method for rapid detection of universal human adenovirus using recombinase polymerase constant temperature amplification (Recombinase Polymerase Amplification, RPA), its special primers and probes and its use . Background technique [0002] There are currently 57 serotypes of adenoviruses that are pathogenic to humans, which can be divided into 7 subgroups, A-G. This virus usually causes mild self-limiting respiratory diseases in healthy people, and in most cases it can be diagnosed clinically. Diagnosed, but sometimes adenoviruses can cause acute respiratory infections in humans, espec...

AI Technical Summary

Problems solved by technology

Human adenovirus can cause regional epidemics or outbreaks, but because the clinical manifestations of its infection are usually non-specific, there is currently no specific treatment. Once it develops into severe pneumonia, the fatality rate is high, especially for types 3 and 7. Types, 11, 21, and 55 are highly contagious, and in places where people gather, such as schools and the military, once they occur, they are likely to cause mass infection and outbreaks

At present, the commonly used molecular biology detection methods include ordinary PCR, real-time fluorescent quantitative PCR, and loop-mediated isothermal amplification. Popularized by the health department, it is not suitable for on-site rapid diagnosis in harsh environments such as field and on-site testing

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