Method for detecting various respiratory viruses and primers and probes thereof

Abstract

The invention belongs to the technical fields of biochips and diagnostic reagents, and discloses a method for detecting various respiratory viruses, and primers and probes thereof. In the invention, nucleotide sequences of 14 respiratory viruses, namely adenovirus, human metapneumovirus, influenza virus A, influenza virus B, respiratory syncytial virus, bocavirus, rhinovirus, coronavirus (HKU1, NL63 and SARS), and parainfluenza virus (type I, type II, type III and type IV) are analyzed, and corresponding reverse transcription primers, PCR primers and specific probes are designed. Specific gene segments are amplified by reverse transcription and multiple asymmetric PCR methods; a fluorescence-coded microsphere group coupled with the virus specific probes and the PCR amplification product are incubated and hybridized by liquid phase chip technology; and finally the Bio-PlexTM200 is used for detection. The detection method has the advantages of high flux, high specificity and sensitivity, stable results and good repeatability, the detection method is easy to operate, and the detection speed is high.

Description

technical field [0001] The invention belongs to the technical field of biochips and diagnostic reagents, in particular to a method for detecting a variety of respiratory viruses and its primers and probes, in particular to the use of specific nucleic acid probes for respiratory viruses combined with multiple polymerases in a liquid phase chip Chain reaction (PCR) technology, a method for the rapid detection of multiple respiratory viruses. Background technique [0002] Respiratory diseases are one of the most common fatal diseases in the world. There are many kinds of pathogenic microorganisms that cause acute respiratory infections, among which viruses account for more than 90%, and the clinical manifestations and symptoms caused by them are similar: such as fever, headache, sore limbs, Sneezing, runny nose, stuffy nose, coughing, sore throat, etc. Common respiratory viruses include adenovirus (AdV), human metapneumovirus (hMPV), influenza virus type A (IfA), influenza vir...

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Problems solved by technology

However, there are more or less defects in these methods: for example, the isolation and culture method is technically difficult, and it usually takes more than 3 to 5 days to obtain the results of etiological diagnosis and drug susceptibility, and it is difficult to carry out etiological diagnosis and drug resistance early in clinical practice. However, when using immunological methods to detect virus-specific antibodies, it is not only necessary to collect double sera in the acute phase and recovery phase of the patient's infection, but there may also be antigen-antibody gaps. Interfering with the detection due to cross-reactions, which is not conducive to the early diagnosis of diseases; in terms of nucleic acid detection methods, PCR, multiplex PCR, RT-PCR methods, and real-time fluorescent PCR methods based on Taqman probe technology have high sensitivity and good specificity. However, most of these methods are based on the detection of a single reaction or a small number of multiple reactions. When testing clinical samples, it takes a lot of labor to detect and exclude dozens of indicators in each sample. and cost

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