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Kits for Detecting 2019 Novel Coronavirus Nucleic Acid

A coronavirus and kit technology, applied in the field of molecular biology, can solve the problems of long detection time, affecting the judgment and prevention and control of the epidemic, and achieve the effect of convenient operation, high sensitivity and high sensitivity detection

Active Publication Date: 2020-12-29
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantage of this method is that the detection takes a long time, and it needs to rely on PCR instrument or expensive real-time quantitative PCR instrument and other supporting equipment for PCR detection
In the detection of new coronavirus, RT-PCR method has many false negative results, which affects the judgment and prevention and control of the epidemic

Method used

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  • Kits for Detecting 2019 Novel Coronavirus Nucleic Acid
  • Kits for Detecting 2019 Novel Coronavirus Nucleic Acid
  • Kits for Detecting 2019 Novel Coronavirus Nucleic Acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034]Designed to detect the crRNA of 2019 novel coronavirus nucleic acid

[0035]Obtain the genome sequence of 2019-nCoV, and find its specific identification region through bioinformatics analysis and comparison.

[0036]The specific operation steps are as follows, search the entire genome of 2019-nCoV and other coronaviruses (SARS-nCoV, MERS-nCoV, HCoV-OC43, HCoV-HKU1, MHV, HCoV-229E, HCoV-NL63, IBV) in the NCBI nucleic acid sequence library Sequence to obtain the whole genome sequence. Search for "TTTN" sequences in the 2019-nCoV sequence, and use these sequences as candidate databases for crRNA targeting sequences.

[0037]Compared with other coronaviruses (SARS-nCoV, MERS-nCoV, HCoV-OC43, HCoV-HKU1, MHV, HCoV-229E, HCoV-NL63, IBV), look for the specificity of the sequence in the candidate database. According to the screening principles, such as the GC content of the sequence, base uniformity, sequence conservation and other parameters, a target site recognition sequence with a size of ...

Embodiment 2

[0039]The kit for detecting the nucleic acid of the 2019 novel coronavirus (the first type) includes: crRNA-1, LbCas12a nuclease (Lba Cas12a (Cpf1), BioLabs), nucleic acid detection probe (the 5'end of the nucleotide is modified with FAM and the 3'end is modified with BHQ-1), the fifth upstream primer, the fifth downstream primer, andDNAAmplification kit (commercially available), water (DEPC treated water) and 1×NEbufferTM 2.1(BioLabs);

[0040]The nucleotide sequence of the nucleic acid detection probe is shown in SEQ ID NO. 3;

[0041]The nucleotide sequence of the fifth upstream primer is shown in SEQ ID NO. 8;

[0042]The nucleotide sequence of the fifth downstream primer is shown in SEQ ID NO.13.

Embodiment 3

[0044]The kit for detecting the nucleic acid of the 2019 novel coronavirus (the second type) includes: crRNA-2 and LbCas12a nuclease (Lba Cas12a (Cpf1), BioLabs), nucleic acid detection probe (the 5'end of the nucleotide is modified with FAM and the 3'end is modified with BHQ-1), the fifth upstream primer, the fifth downstream primer, andDNAAmplification kit (commercially available), water (DEPC treated water) and 1×NEbufferTM 2.1(BioLabs);

[0045]The nucleotide sequence of the nucleic acid detection probe is shown in SEQ ID NO. 3;

[0046]The nucleotide sequence of the fifth upstream primer is shown in SEQ ID NO. 8;

[0047]The nucleotide sequence of the fifth downstream primer is shown in SEQ ID NO.13.

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Abstract

The invention discloses a kit for detecting 2019 novel coronavirus nucleic acid, comprising: crRNA-1 for detecting 2019 novel coronavirus nucleic acid or crRNA-2 for detecting 2019 novel coronavirus nucleic acid, LbCas12a nuclease, nucleic acid detection probe, primer pair and DNA Amplification kit, water and 1×NEbuffer TM 2.1; The kit for detecting 2019 novel coronavirus nucleic acid of the present invention can quickly and accurately detect 2019 novel coronavirus nucleic acid, has good detection specificity, high sensitivity, can realize highly sensitive detection at room temperature, is easy to operate, The advantage of being quick.

Description

Technical field[0001]The invention belongs to the technical field of molecular biology. More specifically, it relates to a kit for detecting nucleic acid of 2019 novel coronavirus (2019-nCoV).Background technique[0002]The 2019 novel coronavirus is a newly discovered new strain of coronavirus that can cause human novel coronavirus pneumonia. The main manifestations are fever, fatigue, and dry cough. In severe cases, it can rapidly progress to acute respiratory distress syndrome and sepsis. Shock, difficult to correct metabolic acidosis and coagulopathy. At present, clinical diagnosis is mainly based on epidemiology, clinical manifestations, and laboratory tests.[0003]At present, in the detection of the 2019 novel coronavirus, the gold standard is still the real-time fluorescent quantitative PCR (Real-time-PCR, RT-PCR) detection technology, which has high sensitivity and specificity. Using real-time fluorescent PCR detection, the real-time fluorescent PCR detection alone generally tak...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2521/107Y02A50/30
Inventor 王涛刘思华黄梦倩徐亚楠王志云
Owner TIANJIN UNIV