RPA method for detecting 21 type human adenovirus, special primer, probe and application thereof

A human adenovirus and probe technology, used in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of long detection time, reliance on expensive instruments, and inability to apply on-site detection.
CN109504801AInactive Publication Date: 2019-03-22李越希
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
李越希
Publication Date
2019-03-22
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a RPA detection method for detecting 21 type human adenovirus, a special primer, a probe and an application thereof in detecting 21 type human adenovirus. The detection method,the special primer and the probe thereof are designed on the basis of 21 type human adenovirus hexon gene conserved sequence, with oligonucleotide sequences shown as SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4. According to the invention, a novel isothermal amplification technology RPA is firstly applied to the detection of 21 type human adenovirus. According to the method, an enzyme reaction processof in vivo DNA replication is simulated; amplification of DNA template relies on a specific combination of enzyme and protein (recombinase, single-stranded binding protein and DNA polymerase); amplification of specific nucleotide sequence is realized under a constant temperature at 37 DEG C; amplified products can be visually identified through a lateral chromatography test strip. The detection method established by the invention has the advantages of high sensitivity, limit of detection reaching up to 102 copies / reaction, high specificity, no cross reaction with other pathogens, high detection speed, only 20min reaction time, simple and convenient operation, and suitability for onsite detection.
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Description

technical field

[0001] The invention discloses an RPA method for detecting type 21 human adenovirus, its special primers and probes and its use, which belong to the field of biotechnology, relate to the molecular biology of type 21 human adenovirus (human adenovirus, HAdV), and relate to a method for detecting type 21 The method and application thereof for human adenovirus, in particular to a method for rapidly detecting type 21 human adenovirus using recombinase polymerase constant temperature amplification (Recombinase Polymerase Amplification, RPA) technology, its special primers and probes and its application. Background technique

[0002] In recent years, there have been occasional outbreaks of HAdV21 infection in densely populated units such as troops and schools. HAdV21 is a highly virulent HAdV, and its main symptom is febrile respiratory infection, which can cause severe pneumonia and even death in severe cases. At present, PCR and q-PCR are the main identification ...

Claims

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