Kit for nucleic acid combined detection of influenza virus A, influenza virus B and respiratory syncytial virus

Abstract

The invention provides a kit for nucleic acid combined detection of the influenza virus A, the influenza virus B and the respiratory syncytial virus. The kit comprises RT-PCR reaction liquid, an RT-PCR enzyme mixture, a respiratory virus primer probe, internal reference, negative contrast, clinical positive contrast and strong positive contrast. The one-step RT-PCR reaction can be directly conducted on the well-extracted respiratory virus nucleic acid, the influenza virus A, the influenza virus B and the respiratory syncytial virus in a sample can be detected in a classified and qualitative mode, the gene of the internal reference serves as the internal contrast, and contamination is prevented through UNG enzymes. The kit is simple in one-step amplification method, short in procedure, easy and convenient to operate, capable of preventing contamination, high in detection result specificity, high in sensitivity, clear in result, high in credibility, and capable of being used for qualitatively authenticating and detecting the influenza virus A, the influenza virus B and the respiratory syncytial virus in a human nose pharynx-mop sample.

Description

Technical field [0001] The present invention is a biotechnology field, involving a biological testing technology, especially involving a combination of type A, B virus and respiratory tract hypotenocyte virus nucleic acid. Background technique [0002] VirusesAssociateDWithRespiratoryinfections refers to the general term that mainly uses the respiratory tract as an invasion of the portal.90-95%of clinical acute respiratory infections are caused by this group of poison.The respiratory virus is mostly a single -chain RNA (SSRNA) virus. It generally has envelope and thorns. It is mainly transmitted through the respiratory tract. It is positioned as the respiratory tract. It can be infected repeatedly and high population.Among them, Influenzavirus and respiratory tract sympathetic virus (RSV) are clinically and important respiratory viruses. [0003] The form of respiratory virus virus is spherical, the genome is a linear, regardless of the section of the single -chain RNA (SSRNA),...

AI Technical Summary

Problems solved by technology

Four domestic companies have developed joint detection reagents for influenza A virus and influenza B virus, all of which have been licensed by the state, but there is no fluorescent PCR detection kit for joint detection of respiratory syncytial virus

[0011] At the same time, in order to avoid false positives in the reaction results, using the anti-pollution characteristics of UNG enzyme, dUTP is used instead of dTTP in the PCR amplification reaction, so that only dUTP is contained in the amplified fragment; Pre-DNA is easily degraded by UNG enzyme: this PCR product containing dUTP is incubated with UNG enzyme, because UDG enzyme can cleave the N-glycosyl bond between uracil base and sugar phosphate backbone, and can be obtained from single-stranded or double-stranded DNA Elimination of dUTP while preventing Taq Elongation by DNA polymerase, thereby losing the ability to be reamplified