Real-time fluorescent nucleic acid constant-temperature amplification and detection kit for human seasonal influenza viruses H1N1 (HuH1N1)

Abstract

The invention discloses a real-time fluorescent nucleic acid constant-temperature amplification and detection kit for human seasonal influenza viruses H1N1 (HuH1N1). The real-time fluorescent nucleic acid constant-temperature amplification and detection kit comprises a capturing probe, a HuH1N1 detection primer T7, a primer nT7, a HuH1N1 detection probe, an M-MLV reverse transcriptase, T7 RNA (ribonucleic acid) polymerase and the like. The kit disclosed by the invention can detect HuH1N1 RNA in a swab, has the characteristics of high specificity, high sensitivity (which is up to 100copies / reaction), low pollution (due to easy degradation of the RNA of amplification products under a natural environment) and quickness in detection ( due to detection normally implemented within 50 minutes), plays an important role in clinical diagnosis of early infection of the human seasonal influenza and has a large application prospect.

Description

technical field [0001] The present invention relates to the biological detection technology of viruses, in particular to the primers used in the real-time fluorescent nucleic acid constant temperature amplification detection of human seasonal influenza virus H1N1 (HuH1N1), which combines specific target capture technology and real-time fluorescent nucleic acid constant temperature amplification detection technology, Probes and related kits. Background technique [0002] Influenza viruses are divided into three types: A, B, and C according to nucleoprotein (NP) and matrix protein (M). The influenza A virus genome consists of 8 negative-strand RNA segments, which are divided into 16 HA subtypes and 9 NA subtypes according to the surface hemagglutinin (HA) and neuraminidase (NA). [0003] Human seasonal influenza virus H1N1 (HuH1N1) belongs to the Orthomyxoviridae Influenza A virus genus, and the typical virus is spherical. Seasonal influenza virus will be popular and spread ...

AI Technical Summary

Problems solved by technology

At present, there are reverse transcriptase-polymerase chain reaction (Reverse Transcriptase-PCR, RT-PCR for short) and real-time fluorescent quantitative polymerase chain reaction (Real Time Fluorescent Quantified PCR, FQ PCR for short), RT-PCR adopts "gene amplification The operation mode of "amplification + amplicon detection" is easy to cause contamination of the amplicon, which often results in false positive or false negative of the experimental results.

Although real-time fluorescence quantitative PCR solves the above problems technically, its sensitivity and accuracy are high, but the operation is complicated and the detection cost is relatively expensive, which hinders its large-scale promotion and use in developing countries.

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