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Nucleic acid reagent, kit, system and method used for detecting avian influenza viruses and/or avian influenza virus drug resistance

A bird flu virus and drug resistance technology, applied in biochemical equipment and methods, specific-purpose bioreactors/fermenters, enzymology/microbiology devices, etc., can solve the problem of limited throughput, inability to obtain, and insufficient detection Sensitivity and other issues, to avoid sample extraction steps, overcome insufficient detection capacity, and improve detection throughput

Active Publication Date: 2019-04-12
北京卓诚惠生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

Commonly used PCR detection requires complex sample processing, cumbersome test procedures and complex result judgments
In addition, highly pathogenic mutations and drug-resistant mutations include point mutations, insertion fragments, deletion fragments, etc., and ordinary TaqMan probes are not sensitive enough to detect point mutations, so they cannot be identified; insertion fragments and deletion fragments will cause TaqMan probes. Unrecognizable, unable to obtain results that can be judged, resulting in false negatives or misjudgments
Can MGB probes recognize SNP sites well, but the overall throughput is limited, and no more than two MGB probes can be added to the reaction system; and MGB probes also cannot recognize insertions and deletions, and cannot obtain available The result of judgment, resulting in false negative or misjudgment
[0007] Conventional nucleic acid detection products can only determine whether it is a known pathogen or detect existing drug resistance sites, but cannot determine new mutations

Method used

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  • Nucleic acid reagent, kit, system and method used for detecting avian influenza viruses and/or avian influenza virus drug resistance
  • Nucleic acid reagent, kit, system and method used for detecting avian influenza viruses and/or avian influenza virus drug resistance
  • Nucleic acid reagent, kit, system and method used for detecting avian influenza viruses and/or avian influenza virus drug resistance

Examples

Experimental program
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Effect test

Embodiment

[0070] 1. Primer and probe synthesis

[0071] Sequence synthesis was performed according to the primer sequences shown in Table 1 and the probe sequences shown in Table 2. In the sequence, Y represents the degenerate base T / C; R represents the degenerate base A / G; W represents the degenerate base A / T; FAM in the probe is 6-carboxyfluorescein, and JOE is 2,7-di Methyl-4,5-dichloro-6-carboxyfluorescein, TAMRA is 6-carboxytetramethylrhodamine, and CY5 is 5H-indocyanine. The brackets in the probe sequences in Table 2 indicate that the t on the left side of the brackets has a fluorescent label, and the contents in the brackets indicate the choice of fluorescent labels.

[0072] Table 1

[0073]

[0074] Table 2

[0075]

[0076] 2. Sample processing

[0077] After the sample is collected with the sampler matched with ParaDNA, it can be amplified directly in the reactor of ParaDNA.

[0078] 3. Construct Hybeacon probe technology detection system

[0079] Polymerase Phire...

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Abstract

The present disclosure relates to a nucleic acid reagent, a kit, a system and a method used for detecting avian influenza viruses and / or avian influenza virus drug resistance. The nucleic acid reagentcomprises primers shown in SEQ ID NO.1-8 and probes shown in SEQ ID NO.11-15, which are stored separately from each other or arbitrarily mixed and stored with each other. The disclosed primers and probes establish the nucleic acid reagent, kit, system and method for detecting the avian influenza viruses and common drug resistance genes carried by the avian influenza viruses. The nucleic acid reagent can realize rapid, comprehensive, sensitive, specific and automatic detection result determination, and significantly improves sensitivity, specificity and simplicity of detecting the avian influenza viruses and drug resistance thereof.

Description

technical field [0001] The present disclosure relates to the field of biotechnology, in particular, to a nucleic acid reagent, kit, system and method for detecting avian influenza virus and / or drug resistance of avian influenza virus. Background technique [0002] The epidemic peak of avian influenza virus in winter and spring every year poses a huge threat to the breeding industry, and can cause human infection and death. The case fatality rate of human infection with highly pathogenic avian influenza virus is as high as 50%. [0003] So far, highly pathogenic avian influenza is caused by H5 and H7 subtype avian influenza virus. Usually, H5 and H7 subtype viruses stably exist in natural hosts in a form of low pathogenicity. However, only a small number of H5 and H7 subtype avian influenza strains are highly pathogenic. These viruses are transmitted from reservoir hosts to poultry by a number of different routes. After several cycles of infection (and possibly adaptive mut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12M1/34
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2600/166C12Q2537/143C12Q2563/107C12Q2527/107C12Q2545/101
Inventor 王晓艳林笑冬王雷张志强
Owner 北京卓诚惠生生物科技股份有限公司
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