Unlock instant, AI-driven research and patent intelligence for your innovation.

Kit for jointly detecting influenza A virus and influenza B virus based on double amplification technology and application of kit

A technology for influenza B virus and influenza A virus, which is applied in the field of kits for joint detection of influenza A and influenza B virus nucleic acids based on dual amplification technology, and can solve the problems of complex RNA extraction process and easy pollution.

Active Publication Date: 2020-05-19
武汉中帜生物科技股份有限公司
View PDF17 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Now commonly used influenza A and B virus nucleic acid detection methods are based on RT-PCR methods, these methods require complex RNA extraction process, special PCR amplification conditions, specialized laboratories and fluorescent quantitative PCR instruments, and extremely difficult during the detection process. prone to pollution

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit for jointly detecting influenza A virus and influenza B virus based on double amplification technology and application of kit
  • Kit for jointly detecting influenza A virus and influenza B virus based on double amplification technology and application of kit
  • Kit for jointly detecting influenza A virus and influenza B virus based on double amplification technology and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] [Example 1] Sensitivity test

[0087] The virus stocks of H1N1 (ATCC No. VR-1469), H3N2 (ATCC No. VR-1680D), and FluB (ATCC No. VR-1735) sourced from ATCC were subjected to serial dilutions to determine the minimum detection limit, and each gradient of virus dilutions Repeat 3 to 5 copies, each for 20 times repeated detection, and the virus level with a positive detection rate of 90% to 95% is taken as the minimum detection limit. The detection results are as follows:

[0088] H1N1 minimum detection limit detection

[0089] Table 1.1 Test data of different titers of H1N1

[0090]

[0091] Table 1.2 Experimental data of the lowest detection limit of H1N1

[0092]

[0093]

[0094] H3N2 minimum detection limit detection

[0095] Table 2.1 Test data of different titers of H3N2

[0096]

[0097] Table 2.2 Experimental data of the lowest detection limit of H3N2

[0098]

[0099]

[0100] FluB minimum detection limit detection

[0101] Table 3.1 Test data of FluB with different titers

[...

Embodiment 2

[0108] [Example 2] Specificity verification

[0109] 1. Test strain

[0110] Different microorganisms are subjected to nucleic acid extraction and detection to verify the specificity of the primer and probe design of the kit of the invention. Related pathogens and titers are as follows:

[0111] Table 4 Specific verification test strain information

[0112]

[0113]

[0114] 2. Test results

[0115] The test results are as follows:

[0116] Table 5 Specificity verification test results

[0117]

[0118]

[0119] 3. Conclusion

[0120] It can be seen from the above data that the test results of the kit of the present invention for these microorganisms are all negative, which proves that there is no cross-reaction between the kit of the present invention and other microorganisms, which reflects the strong specificity of the kit for detecting pathogens.

Embodiment 3

[0121] [Example 3] Verification of pathogen detection ability

[0122] The kit of the present invention is used to detect 28 common influenza virus strains, and the detection ability of the kit for different influenza virus strains is verified. The test results are as follows:

[0123] Table 6 Test results of different virus strains

[0124]

[0125]

[0126] It can be seen from the above results that this kit has a good detection ability for common influenza virus subtypes.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a kit for jointly detecting influenza A virus and influenza B virus based on a double amplification technology and an application of the kit. A collected sample is lysed by cell lysate to release pathogen nucleic acid, and the pathogen nucleic acid fragment is amplified through reverse transcription and transcription processes. The amplified RNA product is added into micropores which coat coating probes, and meanwhile, a specific probe and an amplification probe are added, wherein the coated probe can be combined with one end of the specific probe CES to fix an amplification product RNA; and one end of the specific probe LES is combined on the RNA product, and the other end of the specific probe LES is combined with the amplification probe to realize signal amplification. The amplification probe labeled with multiple biotins is subsequently combined with a streptavidin-HRP enzyme compound, finally an HRP enzyme chemiluminescence substrate is added, and detectionis carried out on a chemiluminescence instrument. According to the invention RNA extraction is not needed, pollution in detection is not easy, sensitivity is high, and specificity is high, so that the kit can be widely used for nucleic acid detection of g influenza A virus and influenza B virus.

Description

Technical field [0001] The invention relates to the technical field of biological detection, in particular to a kit based on double amplification (RNA constant temperature amplification + multi-biotin signal amplification) technology for combined detection of nucleic acid of influenza A and B viruses and its application. Background technique [0002] Influenza viruses include Type A, Type B, and Type C. Type A is the most likely to cause an epidemic, followed by Type B, and Type C rarely causes an epidemic. According to the antigenicity of hemagglutinin (HA) and neuraminidase (NA) proteins in the outer membrane of viral particles, influenza A viruses can currently be divided into 18 H subtypes (H1-H18) and 11 N subtypes ( N1-N11). There have been reports of human infections in subtypes such as H1, H2, H3, H5, H7 and H9. Because the nucleotide sequence encoding HA and (or) NA is prone to mutations, resulting in changes in the epitopes of HA and (or) NA, this antigenic change make...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/682C12Q1/6834C12Q1/70
CPCC12Q1/682C12Q1/6834C12Q1/701C12Q2600/166C12Q2531/119C12Q2547/101C12Q2545/101C12Q2545/113C12Q2563/103C12Q2521/107Y02A50/30
Inventor 李先强姜昕黄永伟陈巨
Owner 武汉中帜生物科技股份有限公司
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com