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Nucleic acid composition, kit and method for detecting 2019 novel coronaviruses

A technology of nucleic acid composition and coronavirus, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/testing, etc., can solve problems such as critical illness and death, achieve simple operation, improve detection sensitivity, and shorten detection the effect of time

Active Publication Date: 2020-04-10
ZHUHAI LIVZON DIAGNOSTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Judging from the current cases, the prognosis of most patients is good, while a few patients are in critical condition, and even co-morbid with other underlying diseases leading to death.

Method used

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  • Nucleic acid composition, kit and method for detecting 2019 novel coronaviruses
  • Nucleic acid composition, kit and method for detecting 2019 novel coronaviruses
  • Nucleic acid composition, kit and method for detecting 2019 novel coronaviruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] The nucleic acid composition for detecting 2019 novel coronavirus provided in this embodiment includes:

[0089] A first nucleic acid combination comprising:

[0090] The first primer pair shown in SEQ ID NO.4 and SEQ ID NO.6, and the first probe shown in SEQ ID NO.5; wherein, the 5' end of the first probe is labeled with a fluorescent reporter group FAM , the 3' end is labeled with a fluorescent quencher group BHQ1;

[0091] A second nucleic acid combination comprising:

[0092] The second primer pair shown in SEQ ID NO.7 and SEQ ID NO.8, and the second probe shown in SEQ ID NO.9; the 5' end of the second probe is labeled with a fluorescent reporter group HEX, 3 The 'end is labeled with a fluorescent quencher group BHQ1;

[0093] And, the third nucleic acid combination, it comprises:

[0094] The third primer pair shown in SEQ ID NO.10 and SEQ ID NO.11, and the third probe shown in SEQ ID NO.12; the 5' end of the third probe is labeled with a fluorescent reporter g...

experiment example 1

[0114] Verify the sensitivity of the nucleic acid composition of Example 1

[0115] The detection method is as follows:

[0116] Two positive plasmids (S gene positive plasmid and N gene positive plasmid) respectively containing the S gene fragment (SEQ ID NO.1) and the N gene fragment (SEQ ID NO.2), the backbone of which is the puc57 vector, the vector map is shown in Figure 37 ) Templates were mixed and diluted to 4.0E+04copies / mL, 4.0E+03copies / mL, 4.0E+02copies / mL, 40copies / mL and 20copies / mL for extraction (for the extraction method, refer to the nucleic acid sample to be tested in Example 1 In other embodiments, the positive plasmid template can be subjected to PCR amplification detection without extraction treatment. In this experimental example, it is extracted, mainly for the purpose of simulating the sample after being extracted by the extraction method used in the present invention. Sensitivity of detection with the kit of the present invention), wherein 20copies / ...

experiment example 2

[0129] Verify the specificity of the nucleic acid composition of Example 1

[0130] Detection of respiratory syncytial virus type A, respiratory virus type B, influenza A virus H1N1, influenza A virus H3N2, influenza A virus H1N1 (2009), influenza B virus Yamagata and influenza B virus Victoria virus and golden yellow grape Bacteria culture, to verify whether there is cross-reaction with the nucleic acid composition of the example.

[0131] The detection method is as follows:

[0132] The above 8 kinds of virus cultures and reference substances (the negative control is normal saline, and the positive control substance contains plasmids of S gene fragments and plasmids of N gene fragments) were extracted according to the extraction method in Example 1, and the extraction volume was 0.5 mL, the elution volume is 60 μL. After the extraction, take 50 μL and add it to the freeze-dried PCR reaction reagent prepared according to the method of Example 1. After mixing, put it on the P...

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Abstract

The invention discloses a nucleic acid composition, a kit and a method for detecting 2019 novel coronaviruses, belonging to the technical field of biology. The nucleic acid composition disclosed by the invention comprises at least one selected from the group consisting of a first nucleic acid combination and a second nucleic acid combination, wherein the first nucleic acid combination comprises nucleic acids as shown in SEQ ID NO.4-6, and the second nucleic acid combination comprises nucleic acids as shown in SEQ ID NO.7-9. When the nucleic acid composition, the kit or the method provided by the invention is used for detecting the 2019 novel coronaviruses, higher sensitivity and higher specificity are achieved.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular, to a nucleic acid composition, kit and method for detecting 2019 novel coronavirus. Background technique [0002] On February 11, 2020, the International Committee on Taxonomy of Viruses (ICTV) announced that the classification of the 2019 novel coronavirus was named Severe Acute Respiratory Syndrome Coronavirus 2 (SARS‑Cov‑2). The disease caused by viral infection is officially named COVID‑19. The 2019 novel coronavirus pneumonia is mainly transmitted by droplets and contact, and the population is generally susceptible. The general symptoms of 2019-nCoV infection are: fever, fatigue, dry cough, and gradual dyspnea. Some patients may have mild onset symptoms or even no obvious fever. Severe symptoms include: acute respiratory distress syndrome, septic shock, difficult-to-correct metabolic acidosis, and coagulation dysfunction. In addition to the above symptoms, there may also...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2563/107Y02A50/30
Inventor 勾宏娜吴燕陈怡欧格邓京
Owner ZHUHAI LIVZON DIAGNOSTICS
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