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Influenza A virus detection kit and detection method thereof

A type of influenza A virus and detection kit technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problems of time-consuming cost, lack of sensitivity and specificity of detection, etc., to achieve The effect of shortening the detection time, avoiding false positive results, and improving the detection rate

Pending Publication Date: 2022-07-01
SHANGHAI ZJ BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In summary, current assays on the market lack sensitivity and specificity, are time consuming and costly

Method used

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  • Influenza A virus detection kit and detection method thereof
  • Influenza A virus detection kit and detection method thereof
  • Influenza A virus detection kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Using the principle of Taqman fluorescence quantitative PCR, specific primers for influenza A virus are designed to amplify the specific nucleic acid sequence of influenza A virus, and Taqman probes are designed for the specific sequence of influenza A virus, located between the upstream and downstream primers. The probe for influenza A virus sequence is labeled with a fluorescent reporter group at the 5' end and a non-fluorescent quencher group at the 3' end. When the probe is intact, the fluorescence energy emitted by the reporter group is absorbed by the quencher group, and the instrument cannot detect the signal. With the progress of PCR, Taq enzyme encounters the probe bound to the template during the chain extension process, and its 5′→3′ exonuclease activity will cut off the probe, and the reporter group is far away from the quencher group, which The energy cannot be absorbed, i.e. a fluorescent signal is generated.

[0112] Ⅰ. Nucleic acid extraction of samples...

Embodiment 2

[0144]Example 2 adopts the same experimental method as Example 1, except that the probe-primer combinations used in the present invention are respectively S1 group, S2 group and S3 group, and the formulas of the probe-primer combination are respectively: (1) S1 group : The third primer set: upstream primer: 0.5 μl / test, downstream primer: 0.5 μl / test; fourth primer set: upstream primer: 0.5 μl / test, downstream primer: 0.5 μl / test, probe: 0.15 μl / test ; (2) S2 group: fifth primer group: upstream primer: 0 μl / test, downstream primer: 0.4 μl / test; sixth primer group: upstream primer: 0.9 μl / test, downstream primer: 0.4 μl / test, probe : 0.15μl / test; (3) S3 group: seventh primer group: upstream primer: 0μl / test, downstream primer: 0.4μl / test; eighth primer group: upstream primer: 0.9μl / test, downstream primer: 0.4μl / test, probe: 0.15 μl / test. The samples are influenza A (untyped) samples, and the results are as follows Figure 10 , Figure 11 , Figure 12 As shown, it shows th...

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Abstract

The invention belongs to the field of virus detection, and particularly relates to application of a sequence as shown in SEQ ID NO.1 as a target to preparation of an influenza A virus detection product. The target sequence disclosed by the invention is relatively good in specificity, and no other similar species exist when the similarity is higher than 80% in a blast result; the conservative property can reach 100% and is far greater than that of other target regions; the sensitivity is higher after gradient dilution. The primer and the probe consider the conservative property in the influenza A virus, can realize full coverage in all known whole genome samples of the influenza A virus, solve the problem of sample missing detection in clinical actual detection, and greatly improve the detection rate of the kit. The design of the specific primer and the probe enables a detected object to be accurate to a seed level, and the false positive rate of detection is greatly reduced due to the advantages of high specificity of a probe technology and high-accuracy quantification of a spectrum technology. And cross reaction is avoided. The detection time is greatly shortened and the operation is simple and convenient. And false positive results caused by pollution of amplification products are avoided.

Description

technical field [0001] The invention belongs to the field of virus detection, in particular to an influenza A virus detection kit and a detection method thereof. Background technique [0002] The most common and most morbid disease in humans is acute respiratory infection (ARIs), of which influenza A is the most common cause of ARIs and lung infections. Influenza A can cause worldwide epidemics, and several cases have occurred in the past 100 years, such as the Spanish H1N1 influenza in 1918, the Asian influenza H2N2 in 1957, the Hong Kong influenza H3N2 in 1968, and the swine influenza H1N1 in 2009. These diseases can infect people of all ages and spread easily and rapidly in the population. According to the official report of the World Health Organization WHO in 2018, influenza A, the 8th deadliest pathogen in the United States, has infected nearly 400 million to 600 million children and 200 million to 50 million adults in the world, with nearly 0.5 to 1 million deaths ea...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2600/16C12Q2531/113C12Q2537/143C12Q2561/101C12Q2563/107
Inventor 邵俊斌张含嫣刘燕舒锋詹晓明
Owner SHANGHAI ZJ BIO TECH
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