Influenza A H1N1/Influenza A Virus Nucleic Acid Dual Fluorescent PCR Detection Kit

A type of influenza A virus detection kit technology, applied in the direction of fluorescence/phosphorescence, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problem of increased detection cost and complicated operation, complex gene sequence determination methods, serum The hysteresis of diagnostic methods can be solved to achieve the effect of rapid and objective test results, strong repeatability, and reduced chances of pollution

Inactive Publication Date: 2011-12-21
JIANGSU BIOPERFECTUS TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005]1) Virus isolation and culture have high technical requirements, are expensive, and time-consuming, and the positive rate of isolation varies from laboratory to laboratory, which cannot meet the requirements of virus epidemic at the same time. The need to handle a large number of samples, currently only used for experimental research
[0006]2) Serological methods cannot process samples with high throughput, and cannot accurately reflect whether they are currently infected or carry the virus
[

Method used

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  • Influenza A H1N1/Influenza A Virus Nucleic Acid Dual Fluorescent PCR Detection Kit
  • Influenza A H1N1/Influenza A Virus Nucleic Acid Dual Fluorescent PCR Detection Kit
  • Influenza A H1N1/Influenza A Virus Nucleic Acid Dual Fluorescent PCR Detection Kit

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1 A H1N1 / Influenza A virus nucleic acid detection kit

[0041] The influenza A H1N1 / influenza A virus nucleic acid double fluorescent quantitative PCR detection kit of the present embodiment includes RNase inhibitor, RT-PCR reaction solution, enzyme mixture, type A H1N1 / influenza A virus double reaction solution, positive control and negative controls,

[0042] Wherein, the RNase inhibitor is DEPC water; the RT-PCR reaction solution includes 10× buffer, MgCl2 and dNTPs;

[0043] Influenza A H1N1 / Influenza A double reaction solution includes the following components:

[0044]Component (1): consists of a pair of primers for detecting influenza A (H1N1) virus and a probe for detecting influenza A (H1N1) virus; wherein, the base sequences of the two primers are SEQ ID No.1 and SEQ ID No. Shown in .2; the base sequence of the probe is shown in SEQ ID No.3, the 5' end of the probe is marked with a fluorescent reporter group, and the 3' end is marked with a fluoresce...

Embodiment 2

[0068] Embodiment 2 Sensitivity test

[0069] The positive reference product is the inactivated virus culture solution, which comes from the Jiangsu Provincial Center for Disease Control and Prevention.

[0070] The negative reference product is RNase-free water. Weigh 1g of DEPC with an electronic balance, add purified water to 1000ml and mix well, then sterilize at 121°C / 20 minutes in a sterilizing pot, mark it, and store it at room temperature.

[0071] The kit of the present invention is used for detection.

[0072] The test results show that the kit of the present invention has good sensitivity, and the CT value changes in a gradient as the concentration decreases ( Figure 4 , Figure 5 ). The test result shows that the kit of the present invention has high sensitivity for the diagnosis of type A H1N1 / type A influenza virus.

[0073]

Embodiment 3

[0074] Embodiment 3 specificity test

[0075] In order to detect the specificity of the A-H1N1 / influenza A virus detection kit of the present invention, the respiratory syncytial virus, human adenovirus, and human parainfluenza virus are detected with the A-H1N1 / influenza A virus detection kit of the present invention.

[0076] The test results show that: the FAM channel only amplifies the influenza A (H1N1) virus ( Figure 6 ), the HEX channel only amplifies influenza A virus ( Figure 7 ). It shows that the detection kit of the present invention can specifically amplify influenza virus without cross-reaction with other viral nucleic acids.

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Abstract

The invention provides a double fluorescent quantitative PCR detection kit of type A H1N1/type A influenza virus nucleic acid, comprising RNase inhibitor, RT-PCR reaction solution, enzyme mixed solution, type A H1N1/type A influenza virus double reaction solution , a positive control and a negative control, wherein said A/H1N1/influenza A double reaction solution includes the following components: Component (1): a pair of primers for detecting A/H1N1 influenza virus and a primer for detecting A Type H1N1 influenza virus probe composition; component (2): composed of a pair of primers for detecting influenza A virus and a probe for detecting influenza A virus; said enzyme mixture includes Taq enzyme and reverse transcriptase . The invention can detect influenza A H1N1 and influenza A virus at the same time, and solves the problem that the existing products can only detect one type of influenza virus in one tube reaction. The invention has the advantages of simple operation, strong repeatability, rapid and objective detection results, etc., and has great application prospects in the field of in vitro diagnosis of influenza virus.

Description

technical field [0001] The invention relates to a dual fluorescent PCR detection kit for simultaneously detecting influenza A H1N1 / influenza A virus nucleic acid, belonging to the field of biotechnology. Background technique [0002] Influenza is a seasonal disease. In temperate regions, influenza prevails throughout the winter. In tropical regions, influenza viruses exist throughout the year, and are more common in rainy seasons. Common influenza A, commonly known as cold, is an acute respiratory infectious disease caused by influenza A virus as the pathogen, and the main mode of transmission is air droplet transmission. The severity of influenza is related to the individual immune status. Generally, only about 50% of infected patients will develop typical clinical symptoms of influenza. Typical symptoms of influenza include sudden fever, dizziness, headache, myalgia, mild systemic symptoms, and may be accompanied by sore throat and cough, nasal congestion, runny nose, ch...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 张旭王国强刘中华魏赵延严浩荣
Owner JIANGSU BIOPERFECTUS TECH CO LTD
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