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Multiple enrichment detection method for influenza A, influenza B and respiratory syncytial virus

A technology of influenza B virus and syncytial virus, applied in the field of molecular biology, can solve the problems of long PCR detection time, high requirement for nucleic acid purity, high automation cost, etc., and achieve improved detection sensitivity, strong specificity, and simple sample processing Effect

Pending Publication Date: 2022-05-03
杭州丹威生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The technical effect of this patented method overcomes previous methods for detecting viruses by combining DNA from both avian (A) and bacteria strains together into one molecule called FluorRed-based probes. This simplifies analysis while improving its ability to identify target sequences quickly compared to other techniques like qPCR or Western blotting.

Problems solved by technology

The technical problem addressed by this patented technology relates to improving the accuracy and efficiency of detecting different types of diseases caused due to both avian-type bacteria or rhinovirus while also reducing their complexity during analysis process compared with existing techniques like polymerase chain reaction (PCR).

Method used

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  • Multiple enrichment detection method for influenza A, influenza B and respiratory syncytial virus
  • Multiple enrichment detection method for influenza A, influenza B and respiratory syncytial virus
  • Multiple enrichment detection method for influenza A, influenza B and respiratory syncytial virus

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Embodiment 1

[0035] The development of embodiment 1 influenza A virus, influenza B virus and respiratory syncytial virus detection reagent

[0036] 1. Design of primers and probes

[0037] According to literature reports, the conserved region corresponding to influenza A virus, influenza B virus and respiratory syncytial virus was selected as the target gene of the kit of the present invention, and 2 pairs of primers and corresponding probes were designed for each virus. And according to the experimental results, considering the detection rate, amplification line type, amplification efficiency and other aspects, a primer combination was selected for each virus. The primer pairs and probe sequences of each virus finally selected are shown in Table 2.

[0038] Table 2

[0039]

[0040] The design of internal standard primer probe of the present invention:

[0041] The present invention introduces the human gene as the internal standard, comprehensively considers the inhibition caused b...

Embodiment 2

[0056] The detection of embodiment 2 type A, type B influenza virus or respiratory syncytial virus positive sample

[0057] A multiple enrichment detection method for influenza A, influenza B and respiratory syncytial virus, the flowchart is shown in figure 1 , the specific process is:

[0058] 1. Method

[0059] 1. Prepare combined detection reagents for influenza A / B virus and respiratory syncytial virus nucleic acid (extraction-free fluorescent PCR method).

[0060] (1) Preparation of PCR reaction solution

[0061] raw material name Volume per serving (μL) Enzyme Mix 1.6 Amplification reaction solution 1 4 Amplification reaction solution 2 9.4 sample 5

[0062] (2) Aliquot PCR reaction solution

[0063] Aliquot the PCR reaction solution into PCR tubes at 15 μL / reaction.

[0064] (3) Preparation of immunomagnetic beads: three antibodies of influenza A, influenza B and respiratory syncytial virus were coupled to carboxyl-modified ...

Embodiment 3

[0092] A kind of primer probe combination that detects influenza A, influenza B and respiratory tract syncytial virus, this combination is made up of the primer probe shown in table 2, and the internal standard primer probe shown in table 3. The combination is used for the detection of influenza A, B or positive samples of respiratory syncytial virus.

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Abstract

The invention relates to the field of molecular biology, in particular to a multiple enrichment detection method for influenza A, influenza B and respiratory syncytial virus. The method comprises the following steps: mixing a respiratory tract sample with immunomagnetic beads respectively coupled with an influenza A virus antibody, an influenza B virus antibody and a respiratory syncytial virus antibody, carrying out mixed incubation at a certain temperature, separating an immunomagnetic bead-virus compound by using a magnetic tool, resuspending the compound in a salt ion buffer solution, and carrying out freeze-drying to obtain the immunomagnetic bead-virus composite. And heating and cracking the resuspended immunomagnetic bead-virus compound, separating the magnetic beads by using a magnetic tool, and directly detecting the cracking product (liquid part) by using a fluorescent PCR detection reagent.

Description

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Claims

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Application Information

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Owner 杭州丹威生物科技有限公司
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