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Reagent kit for influenza a H3N2 type fluorescent PCR diagnosis and use method thereof

A diagnostic kit, H3N2 technology, applied in the field of medical detection, can solve the problems of poor specificity and low sensitivity, and achieve the effects of excellent detection sensitivity, good treatment, and excellent RNA extraction rate

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

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a kind of influenza A H3N2 fluorescent PCR diagnostic kit and its use method, to solve the problems of low sensitivity and poor specificity of existing detection methods

Method used

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  • Reagent kit for influenza a H3N2 type fluorescent PCR diagnosis and use method thereof
  • Reagent kit for influenza a H3N2 type fluorescent PCR diagnosis and use method thereof
  • Reagent kit for influenza a H3N2 type fluorescent PCR diagnosis and use method thereof

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Embodiment 1

[0030] The present invention provides an influenza A H3N2 fluorescent PCR diagnostic kit. The influenza A H3N2 fluorescent PCR diagnostic kit includes an RNA extraction solution and a PCR reaction solution, wherein the RNA extraction solution includes:

[0031] (1) RNA extraction solution I: composed of sodium dodecyl sulfate with a mass / volume ratio of 0.2% to 1.0%, triton with a volume / volume ratio of 1.0% to 4.0%, 0.2mol / L to 1.0mol / L of guanidine isothiocyanate and 100~400μg / ml magnetic beads;

[0032] (2) RNA extraction solution II: 100-300 mmol / L 4-hydroxyethylpiperazineethanesulfonic acid and 100-300 mmol / L, pH6.5±0.2 sodium chloride;

[0033] (3) RNA extraction solution III: volume / volume ratio of 0.1% to 1.0% triton and 100 to 300 mmol / L of sodium chloride;

[0034] (4) RNA extraction solution IV: mineral oil;

[0035] The PCR reaction solution includes: 0.2 μmol / L~0.4 μmol / L of upstream primers and downstream primers for target polynucleotide amplification and 0.2...

Embodiment 2

[0058] see figure 1 , shows the flow chart of the use method of the influenza A H3N2 fluorescent PCR diagnostic kit provided by the present invention.

[0059] This embodiment provides the use method of the influenza A H3N2 fluorescent PCR diagnostic kit described in the above embodiment 1 as follows:

[0060] S101: Take 1200 μl~1ml / human portion of RNA extraction solution I and 1 μl / human portion of the internal standard, mix them uniformly to form RNA extraction reagent I-mix, and use it after instantaneous centrifugation;

[0061] S102: According to the number of samples to be tested, negative controls and positive controls, press 39 μl / person of PCR reaction solution and 1 μl / person of Mn 2+ The ratio of the solution is to take the corresponding amount of the PCR reaction solution and the enzyme mixture, fully mix it into a PCR-mix, and centrifuge it briefly for later use;

[0062] S103: Add 200μl~1ml of RNA extraction solution 1-mix to each tube, then add 100μl~1ml of the...

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Abstract

The invention provides a reagent kit for influenza a H3N2 type fluorescent PCR diagnosis. The reagent kit comprises RNA extraction solutions and a PCR reaction solution. The RNA extraction solutions comprise the RNA extraction solution I, the RNA extraction solution II, the RNA extraction solution III and the RNA extraction solution IV. The RNA extraction solution I is prepared from lauryl sodium sulfate, triton, guanidine thiocyanate and magnetic beads. The RNA extraction solution II is prepared from 4-(2-hydroxyerhyl) piperazine-1-erhaesulfonic acid and sodium chloride. The RNA extraction solution III is prepared from triton and sodium chloride. The RNA extraction solution IV is prepared from mineral oil. The PCR reaction solution comprises an upstream primer, a downstream primer and a probe, wherein the upstream primer and the downstream primer are used for target polynucleotide amplification, and the probe is used for target polynucleotide detection. The reagent kit is an influenza a H3N2 type fluorescent PCR diagnosis product excellent in RNA extraction rate and detection sensitivity. Typing detection can be carried out on influenza a viruses in throat swabs and various samples to judge whether patients are infected with the influenza a H3N2 type or not and assist doctors in making better diagnosis and more precise treatment.

Description

technical field [0001] The invention relates to the technical field of medical detection, and more particularly, to an influenza A H3N2 fluorescent PCR diagnostic kit and a method for using the same. Background technique [0002] Influenza virus is a single-stranded negative-strand virus belonging to the Orthomyxoviridae family. It is spherical or filamentous. The diameter virus consists of three layers. Liposomes are composed of a layer of membrane protein (M). The M protein is antigenically stable and type-specific. The outer layer is a radial protrusion composed of two different glycoproteins, namely hemagglutinin (HA) and neuraminidase. (NA). According to the difference with the antigen, it can be divided into many subtypes, H can be divided into 16 subtypes, and N has 9 subtypes (N1 ~ N9). Among them, H1N1, H2N2, H3N2 and H1N1 new type A, which was popular in 2009, mainly infect humans, and the natural hosts of other subtypes are mainly a variety of birds and animals....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/701C12Q1/686C12Q2545/101C12Q2545/113C12Q2521/531
Inventor 戴立忠范文洲陈晓亮
Owner SANSURE BIOTECH INC
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