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Composition, kit and method for detecting coronaviruses

A technology of coronavirus and composition, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve the problem of no specific treatment for diseases and achieve good stability

Pending Publication Date: 2020-11-27
GENEMIND BIOSCIENCES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no specific treatment for the disease caused by the 2019 novel coronavirus, but many symptoms can be treated with adjuvant care and supportive treatment

Method used

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  • Composition, kit and method for detecting coronaviruses
  • Composition, kit and method for detecting coronaviruses
  • Composition, kit and method for detecting coronaviruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The preparation of embodiment 1 kit

[0036] The preparation method of the kit is exemplified below.

[0037]The kit includes tube 1, which is a primer mixture for amplifying the novel coronavirus (SARS-CoV-2) ORF1ab and N genes. The primer mixture consists of 4 primer sequences, and the specific primer sequences are shown in Table 1. In one case, tube 1 also contains a probe mixture, which is respectively the probes of the novel coronavirus (SARS-CoV-2) ORF1ab and the N gene conserved region, respectively labeled with different fluorescent molecules, in this implementation In the example, the probe of ORF1ab gene marks FAM, and the N gene marks HEX. The specific sequences of the probes are shown in Table 1. Tube 1 of each primer and each probe was used in a concentration range of 8-15uM, preferably 10uM. The novel coronavirus (SARS-CoV-2) ORF1ab and N genes can be effectively amplified by using the kit containing tube 1, and the copy number of the novel coronavirus ...

Embodiment 2

[0084] Embodiment 2 specific detection

[0085] With influenza A virus H1N1 (pdm09), influenza A virus H3N2, influenza B virus Victoria, influenza B virus (Yamagata), influenza C virus, respiratory syncytial virus type A, respiratory syncytial virus type B, Human bocavirus, human parainfluenza virus, human metapneumovirus, rotavirus / enterovirus, adenovirus, human coronavirus (HKU1, OC43, 229E and NL63), Middle East respiratory syndrome coronavirus and other viral nucleic acids as templates, The fluorescent quantitative PCR detection was carried out using the above kit, and the reaction system configuration during detection was set according to Table 3, and the reaction process was set according to Table 4. The test results showed that the above virus test results were all negative.

Embodiment 3

[0086] Application of embodiment 3 kit

[0087] The kit prepared in Example 1 is used for detection of ORF1ab and / or N gene amplification and non-diagnostic purposes containing novel coronavirus (SARS-CoV-2), for example, whether ORF1ab and / or N gene are contained in the plasmid, pure Detection of virus sample concentration, detection of fake virus amount, etc.

[0088] During the detection process, calculate the total number of reactions (N) according to the number of samples to be tested, making sure to include the negative control (1 reaction), positive control (1 reaction) and each sample. According to the number of reactions (N), prepare the PCR reaction system according to Table 6.

[0089] Table 6

[0090]

[0091] In this example, the 2019-nCOV pseudovirus was used as the test sample. The pseudovirus was purchased from Shanghai Yisheng Biotechnology Co., Ltd., the article number is 11900ES03, and the 2019-nCOV pseudovirus is the integration of some genes of the COVI...

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PUM

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Abstract

The present invention provides a composition for detecting coronavirus. The composition comprises sequences represented by Seq ID: 1 and Seq ID: 2 and / or sequences represented by Seq ID: 3 and Seq ID:4, and the composition further comprises sequences represented by Seq ID: 5 and Seq ID: 6. By utilizing the composition provided by the invention, effective amplification and fluorescent quantitativePCR detection can be carried out on an ORF1ab gene and / or an N gene of novel coronavirus, the detection steps are simple, the pollution risk is reduced, and the target gene can be rapidly, efficiently and accurately detected. The invention also provides a kit for detecting the coronavirus and a method for detecting the coronavirus for non-diagnostic purposes. The method has good stability, sensitivity and specificity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, the invention relates to a composition, a kit for detecting coronaviruses and a method for detecting coronaviruses for non-diagnostic purposes. Background technique [0002] Coronaviruses are non-segmented single-stranded positive-sense RNA viruses belonging to the order Nidovirales (Nidovirales) Coronaviridae (Orthocoronavirinae), which are divided into subfamilies according to serotype and genome characteristics The four genera α, β, γ, and δ can infect many animal species, including humans, bats, dogs, pigs, mice, birds, cattle, whales, horses, goats, monkeys, etc. There are 6 coronaviruses known to infect humans, including 229E and NL63 of the genus α, OC43 and HKU1 of the genus β, Middle East respiratory syndrome-associated coronavirus (MERSr-CoV) and severe acute respiratory syndrome-associated coronavirus (SARSr-CoV). CoV). The new coronavirus that broke out in 2019 belongs to...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2563/107C12Q2531/113C12Q2545/113
Inventor 张萌刘磊李改玲李妍冯燕
Owner GENEMIND BIOSCIENCES CO LTD