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Universal coronavirus PCR primers and applications thereof

A general-purpose technology for coronaviruses, applied in the detection/inspection of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve the problems of low cost and high cost, and achieve low cost, high specificity, and easy operation simple effect

Pending Publication Date: 2020-12-11
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional identification methods mainly use fluorescent quantitative primers and probes and ELASA antibody detection, etc., which are highly technical and costly. Therefore, there is an urgent need to detect coronaviruses with low cost, low requirements for instruments and technicians, and simple operation. Quickly and accurately identify whether animals are carrying or infected with coronavirus, so as to play an early warning role in the prevention of coronavirus

Method used

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  • Universal coronavirus PCR primers and applications thereof

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Effect test

Embodiment 1

[0018] 1. A general-purpose coronavirus PCR primer: upstream primer P24F: 5'-AACTCAAGCCTTACCGCAG

[0019] A-3', downstream primer P24R: 5'-ATAGCCCATCTGCCTTGTGT-3'.

[0020] 2. Samples to be tested: for live animals, use sterile cotton swabs to collect animal throat swabs, anal swabs, feces, etc.; for dead individuals, dissect internal organs for use. The samples identified in this example are the feces of a healthy Malayan pangolin (No. 1), the lymph nodes of a dead Malayan pangolin (No. 2), the anal swabs of 2 horseshoe bat individuals (No. 3 and 4), 2 throat swabs from mouse snakes (No. 5 and 6), and 1 throat swab from bamboo rat (No. 7).

[0021] 3. cDNA acquisition: Use the virus extraction kit (QIAamp® Viral RNA Mini Kit) to extract the sample RNA virus, and perform reverse transcription to synthesize single-stranded cDNA according to the instructions of the reverse transcription kit (SuperScript III First-Strand Synthesis System);

[0022] 4. PCR amplification: using c...

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Abstract

The present invention provides universal coronavirus PCR primers and applications thereof. The primers are as follows: an upstream primer P24F: 5'-AACTCAAGCCTTACCGCAGA-3'; and a downstream primer P24R: 5'-ATAGCCCATCTGCCTTGTGT-3'. By utilizing the provided universal coronavirus PCR primers, results can be judged only by using common PCR reagents and an electrophoresis method, operation is simple, and cost is low. Through PCR verification of throat swabs, anus swabs, excrement and lymph node samples of various animals, the universal coronavirus PCR primers have strong specificity, can also be practically applied to identification of coronaviruses carried or infected by the various animals, have characteristics of high specificity, high sensitivity, low cost and simple operation, and can be applied to specific detection of the coronaviruses in animal samples.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a general-purpose coronavirus PCR primer and an application thereof. Background technique [0002] Since the 1970s, there have been more than 40 new infectious diseases in the world, and more than 80% of them originated from wild animals. As an important human public health problem, zoonotic diseases have been paid more and more attention by countries all over the world. In recent years, with the in-depth research on the traceability of infectious diseases, it has been found that wild animal diseases are directly related to human health. [0003] Coronavirus (Coronavirus) is a kind of virus that can cause digestive tract and respiratory diseases in animals and humans. They are a large family. More than 30 kinds have been found so far. It is the virus with the largest genome among RNA viruses, with a genome of about 30,000 cores. nucleotide size. The diameter is 60-140 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2565/125Y02A50/30
Inventor 李林妙陈金平周佳滨刘苹黄文忠
Owner INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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