Primer probe set, kit and detection method for multiple detection of human parainfluenza virus subtypes based on fluorescence RMA method

Abstract

The invention belongs to a detection method for human parainfluenza virus subtypes, and particularly relates to a primer probe group, kit and detection method for multiple detection of human parainfluenza virus subtypes based on a fluorescence RMA method. The kit comprises a detection tube containing an amplification reaction reagent, a buffer solution, magnesium acetate, a standard positive plasmid and sterile double distilled water, wherein the amplification reaction reagent comprises a primer and probe of HPIV1-3, M-MLV reverse transcriptase, escherichia coli RecA protein, UvsY protein, single-stranded binding protein GP32, Bst polymerase, exonuclease III, polyethylene oxide, trehalose, mannitol, ATP, dNTPs, creatine kinase and phosphocreatine; and the standard positive plasmid is a recombinant plasmid obtained by connecting an HN gene segment of HPIV1 with a pMD18-T vector.

Description

technical field [0001] The present application belongs to a detection method for human parainfluenza virus subtypes, specifically a primer probe set, a kit and a detection method for multiple detection of human parainfluenza virus subtypes based on fluorescence RMA method. Background technique [0002] Human parainfluenza virus (HPIV) is a common and easily overlooked pathogen of respiratory tract infection, which mainly causes severe respiratory tract infection in infants, children and immunosuppressed people. In adult patients, parainfluenza virus infection can also lead to a series of respiratory diseases, including upper respiratory tract infection, acute exacerbation of chronic diseases, pneumonia, etc. The clinical manifestations are diverse, and the etiological manifestations are similar to those of other common respiratory infections. In addition, the lack of serological protection also promotes recurrent infection with parainfluenza virus. Human parainfluenza virus...

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Problems solved by technology

[0003] At present, there are many methods for detecting parainfluenza virus, but all of them have deficiencies. For example, the virus isolation and culture method takes a long time and has many false negatives, which cannot be used as a rapid diagnosis method for parainfluenza virus; the detection speed of direct immunofluorescence detection method is fast, but Sensitivity is not as good as that of tissue culture, and rich experience is needed to judge the results; enzyme-linked immunoassay detects virus-specific antibodies but can only detect one virus at a time; fluorescent quantitative PCR method has complex operating procedures, requires sophisticated instruments, and takes a long time to detect. It is not conducive to on-site testing in non-laboratory environments and the promotion and application of grassroots laboratories

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