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Qualitative detection kit for influenza A virus subtype H1N1 nucleic acid

A technology for influenza virus and qualitative detection, applied in the field of diagnosis, can solve problems such as complicated steps and operations

Inactive Publication Date: 2016-02-24
FAPON BIOTECH INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional nucleic acid qualitative detection kits for influenza A (H1N1) viruses usually need to extract purified nucleic acid molecules (H1N1-RNA) from samples, and the steps for extracting nucleic acids from samples are complicated.

Method used

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  • Qualitative detection kit for influenza A virus subtype H1N1 nucleic acid
  • Qualitative detection kit for influenza A virus subtype H1N1 nucleic acid
  • Qualitative detection kit for influenza A virus subtype H1N1 nucleic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Detection of linear templates after the reaction system in Example 1

[0060] The optimization of the detection system for H1N1-1 virus nucleic acid-free nucleic acid extraction is shown in Table 1 below:

[0061] Table 1 Reaction solution configuration for sample detection of H1N1-1 virus nucleic acid

[0062]

[0063] Among them, the optimized concentration of each component in the sample treatment agent is respectively: the concentration of guanidine isothiocyanate is 2mol / L, the concentration of sodium citrate (PH7.0) is 12mmol / L, sodium lauryl creatine The concentration of β-mercaptoethanol is 0.2%, the concentration of β-mercaptoethanol is 0.05mol / L, the concentration of proteinase K is 1mg / mL, and the concentration of H1N1 internal standard is 5×10 3 copies / mL;

[0064] Among them, the optimized concentrations of each component in the 5×RNA one-step reaction buffer are:

[0065] The final concentration of Tris-HCl in the PCR reaction buffer solution is 5mmo...

Embodiment 2

[0083] Preparation and detection of the quality control product of embodiment 2 kit

[0084] H1N1 positive quality control products include strong positive quality control products and borderline positive quality control products, and strong positive quality control products contain about 5×10 5 Copies / mL of non-infectious H1N1RNA in vitro transcripts, the critical positive quality control product contains about 5×10 2 Copies / mL of non-infectious H1N1 RNA in vitro transcripts; negative quality control substance is normal saline.

[0085] The optimized reaction system and reaction conditions were used to detect the prepared quality control products, and the results were as follows: image 3 As shown, the detection of strong positive quality control, critical positive quality control and negative quality control conforms to the product standard of the kit and can be used for the production and quality control of the kit.

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Abstract

The invention discloses a qualitative detection kit for influenza A virus subtype H1N1 nucleic acid. The kit comprises a sample treating agent, an upstream primer HIV-F, a downstream primer HIV-R, a Taqman probe HIV-P and RNA one-step-process reaction buffer. The sample treating agent is prepared from guanidinium isothiocyanate, sodium citrate, sodium dodecyl creatine, beta-mercaptoethanol and proteinase K. The upstream primer HIV-F is used for target nucleotide amplification, the downstream primer HIV-R is used for target nucleotide amplification, the Taqman probe HIV-P is used for target nucleotide detection, and a fluorophore and a fluorescence quencher are combined at the two ends of the Taqman probe HIV-P respectively. In the detection process of the qualitative detection kit for influenza A virus subtype H1N1 nucleic acid, a detection sample and the sample treating agent are directly mixed and then can be used for following detection. Compared with a traditional qualitative detection kit for HIV viruses, the step for extracting purified nucleic acid molecules from the sample is omitted through the qualitative detection kit for influenza A virus subtype H1N1 nucleic acid, and therefore more time is saved during operation and operation is easy.

Description

technical field [0001] The invention relates to the technical field of diagnosis, in particular to a qualitative detection kit for influenza A (H1N1) virus nucleic acid. Background technique [0002] Influenza A (H1N1) virus is a type A influenza virus that carries H1N1 subtype swine influenza virus strains, including ribonucleic acid gene fragments of three influenza viruses: avian influenza, swine influenza and human influenza, as well as Asian swine influenza and African swine influenza Influenza virus characteristics. The crowd is generally susceptible to influenza A (H1N1) virus and can be transmitted from person to person. The early symptoms of human infection are similar to common influenza, including fever, cough, sore throat, body pain, headache, chills and fatigue. Diarrhea or vomiting, muscle pain or tiredness, red eyes, etc. may also occur. Beginning in 2009, influenza A (H1N1) became widespread worldwide. In August 2010, the World Health Organization announce...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q2561/101C12Q2561/113C12Q2545/101
Inventor 何志强范凌云李泓彦
Owner FAPON BIOTECH INC
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