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Kit for simultaneously detecting nucleic acids of seven respiratory pathogens and application thereof

A respiratory and pathogenic technology, which is applied in the field of kits for joint detection of seven respiratory pathogenic nucleic acids based on double amplification technology, which can solve problems such as easy pollution

Active Publication Date: 2020-04-07
武汉中帜生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The commonly used nucleic acid detection methods for respiratory pathogens are based on RT-PCR methods. These methods require complex RNA extraction processes, special PCR amplification conditions, specialized laboratories and fluorescent quantitative PCR instruments, and are very prone to occurrence during the detection process. Pollution

Method used

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  • Kit for simultaneously detecting nucleic acids of seven respiratory pathogens and application thereof
  • Kit for simultaneously detecting nucleic acids of seven respiratory pathogens and application thereof
  • Kit for simultaneously detecting nucleic acids of seven respiratory pathogens and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0152] [Example 1] Sensitivity Test

[0153] The minimum detection limit is determined by gradient dilution of the virus stock solution derived from ATCC. Each gradient virus dilution is repeated 3 to 5 times, and each copy is repeated 20 times. The positive detection rate of 90% to 95% will be determined. virus level as the minimum detection limit,

[0154] The pathogen information is as follows:

[0155] Table 1 Pathogen information

[0156] Subtype ATCC number Corresponding unit H1N1 VR-1469 8.89×10 7 TCID 50 / mL

H3N2 VR-1680D 3.1x 10 6 TCID 50 / mL

FluB VR-1735 2.81×10 7 TCID 50 / mL

RSVA VR-26 1.58×10 7 TCID 50 / mL

RSVB VR-1580 8.89×10 5 TCID 50 / mL

PIV1 VR-94 1.58×10 4 TCID 50 / mL

PIV2 VR-92 2.8×10 6 TCID 50 / mL

PIV3 VR93 1.58×10 7 TCID 50 / mL

AdVB VR-3 1.58×10 8 TCID 50 / mL

AdVE VR-1572 2.8×10 6 TCID 50 / mL

...

Embodiment 2

[0231] [Example 2] Specificity Verification

[0232] 1. Test Strains

[0233] After extracting nucleic acids from different microorganisms, they are tested to verify the specificity of the design of the primers and probes of the kit of the present invention. Relevant pathogens and titers are as follows:

[0234] Table 27 Specificity verification test strain information

[0235]

[0236] 2. Test results

[0237] The test results are as follows:

[0238] Table 28 Specificity Verification Test Results

[0239]

[0240]

[0241] 3. Conclusion

[0242] As can be seen from the above data, the detection 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, reflecting the strong specificity of the kit for detecting pathogens.

Embodiment 3

[0243] [Example 3] Validation of pathogen detection capability

[0244] Use the kit of the present invention to detect 29 strains of pathogens, and verify the ability of the kit to detect different strains of pathogens. The test results are as follows:

[0245] Table 29 Detection results of different pathogen strains

[0246]

[0247]

[0248] It can be seen from the above results that the kit has good detection ability for different pathogen strains.

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PUM

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Abstract

The invention discloses a kit for simultaneously detecting nucleic acids of seven respiratory pathogens and application of the kit. The kit is based on a double amplification technology (RNA isothermal amplification and multi-biotin signal amplification), and can detect H1N1 / H3N2 type influenza A viruses, influenza B viruses, respiratory syncytial viruses, 1 / 2 / 3 type human parainfluenza viruses, B / E type adenoviruses, mycoplasma pneumoniae and chlamydia pneumoniae. The kit of the invention does not need RNA extraction, and is not prone to pollution, high in sensitivity and good in specificityin the process of detection, and can be widely applied to detection of the nucleic acids of the above seven respiratory pathogens.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a kit for jointly detecting nucleic acids of seven respiratory pathogens based on double amplification (RNA constant temperature amplification + multi-biotin signal amplification) technology and an application thereof. Background technique [0002] Influenza virus (FLU) is an enveloped, single-stranded RNA virus with a segmented genome belonging to the Orthomyxoviridae family. According to the antigenicity of the nucleoprotein and the M1 protein on the inner side of the envelope, influenza viruses are divided into three types: A, B, and C. Influenza A virus is divided into several subtypes according to the antigenicity of the two spikes on the surface of the envelope, hemagglutinin (HA) and neuraminidase (NA), currently including subtypes H1-H16 and subtypes N1-N9. The subtypes of influenza viruses are named by listing HA and NA together. H1N1 and H3N2 are relative...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/689C12Q1/6844C12Q1/682C12Q1/04C12R1/93C12R1/35C12R1/01
CPCC12Q1/682C12Q1/6844C12Q1/689C12Q1/701C12Q2600/166C12Q2521/107C12Q2521/327C12Q2545/101C12Q2563/131C12Q2563/125C12Q2563/103
Inventor 李先强姜昕厉洁黄永伟陈巨龚丽君
Owner 武汉中帜生物科技股份有限公司
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