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Dual detection kit for 2019 novel corona virus

A technology of coronavirus and kit, which is applied in the field of primer probe combination and kit for detection of new coronavirus 2019-nCoV, which can solve the problems of CT cross-infection, lack of supporting equipment in epidemic areas, expensive CT equipment, etc.

Active Publication Date: 2020-06-19
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 1. CT is easy to cause cross-infection, unless completely enclosed separate equipment is used, and disinfection must be carried out at all times. For most of the epidemic areas, the equipment and manpower cannot keep up with the consumption;
[0005] 2. There is also the possibility of misdiagnosis when CT is positive. For patients with common influenza, CT images also have ground glass opacity;
[0006] 3. CT equipment is expensive, and remote and underdeveloped epidemic areas cannot be equipped with relevant equipment

Method used

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  • Dual detection kit for 2019 novel corona virus
  • Dual detection kit for 2019 novel corona virus
  • Dual detection kit for 2019 novel corona virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0115] Example 1 ORF1ab gene and N gene double detection kit detection limit test

[0116] Dilute the ORF1ab gene and N gene pseudoviruses with determined values ​​as initial samples to a concentration of 10 5 copies / ml, serially diluted to 10 4 、10 3 , 500, 250, 100 copies / ml, each concentration sample was added to the final concentration of 10 4 Copies / ml of the plasmid bacteria containing the amplified fragment of the internal standard is used as the sample to be tested, and the sensitivity of the dual detection reagent is tested.

[0117] Test result reference Figure 1-3 :

[0118] figure 1 : ORF1ab gene detection limit;

[0119] figure 2 : N gene detection limit;

[0120] image 3 : internal standard test result.

[0121] The test results showed that the lowest detectable concentration was 250 copies / ml for the detection of positive quality control substances of ORF1ab gene and N gene at different concentrations.

Embodiment 2

[0122] Example 2 ORF1ab gene and N gene double detection kit specificity test

[0123] Use normal saline, SARS virus, rhinovirus, adenovirus, influenza A virus, influenza B virus, parainfluenza virus, coronavirus OC43, respiratory syncytial virus, human coronavirus NL63, human coronavirus HKU1 as specificity Reference product, to test the specificity of ORF1ab gene and N gene double detection kit.

[0124] Test result reference Figure 4-5 :

[0125] Figure 4 : ORF1ab gene and N gene specific test results;

[0126] Figure 5 : ORF1ab gene and N gene-specific detection internal standard test results.

[0127] The test results showed that the test-specific reference products (normal saline, SARS virus, rhinovirus, adenovirus, influenza A virus, influenza B virus, parainfluenza virus, coronavirus OC43, respiratory syncytial virus, human coronavirus NL63 , human coronavirus HKU1), the test results were all negative, and the internal standard quality controls were all positi...

Embodiment 3

[0128] Example 3 ORF1ab gene and N gene double detection kit precision test

[0129] Dilute ORF1ab gene and N gene pseudovirus to 10 5 and 10 3 Copies / ml was used as a precision reference product, each repeated 10 times, and the coefficient of variation of each concentration precision reference product was calculated.

[0130] Test result reference Figure 6-7 :

[0131] Image 6 : ORF1ab gene precision test results

[0132] Figure 7 : N gene precision test result

[0133] The coefficients of variation of the tested ORF1ab gene pseudovirus high concentration and low concentration precision reference products were 0.55% and 0.65% respectively; the variation coefficients of the N gene high concentration and low concentration precision reference products were 0.44% and 0.96% respectively; The coefficients of variation of precision reference products with different concentrations of genetic pseudoviruses were all less than 5%.

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Abstract

The invention provides a dual detection kit for a 2019 novel corona virus (2019-nCoV). Specifically, the invention discloses a kit and method for multiple detection of 2019-nCoV nucleic acid. The kitcan simultaneously detect two nucleic acid targets of the 2019-nCoV, has extremely high sensitivity and specificity, and significantly improves the accuracy of virus identification.

Description

technical field [0001] The invention belongs to the field of biotechnology and molecular diagnosis, in particular, the invention relates to a combination of primers and probes and a kit for detecting novel coronavirus 2019-nCoV. Background technique [0002] The new coronavirus (2019 novel corona virus, 2019-n CoV, SARS-CoV-2) belongs to the β-coronavirus genus, has an envelope, and the particles are round or oval, often pleomorphic, with a diameter of 60-140nm. The S protein is one of the main proteins of the virus, and its encoded gene is used for virus typing. The molecular mechanism of the interaction between the S-protein and human ACE2 is used to infect human respiratory epithelial cells. The common sign of people infected with this coronavirus is fever. , cough, shortness of breath and difficulty breathing. Severe infections can lead to pneumonia, severe malformed respiratory syndrome, kidney failure, and even death. This new type of coronavirus is highly infectious...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12N15/11C12Q2600/16C12Q2537/143Y02A50/30C12N2770/20011
Inventor 蒋析文范建彭海龙彭荣俞
Owner DAAN GENE CO LTD
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