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Fish probe set for detecting novel coronavirus sars-cov-2 and its preparation method and use

A coronavirus, sars-cov-2 technology, applied in the field of FISH probe sets for the detection of the new coronavirus SARS-CoV-2, can solve the problem of missed detection rate, inability to quantify and locate the virus, and improve the lack of NCP detection ability , the effect of high sensitivity and short detection period

Active Publication Date: 2020-09-01
SHANGHAI GENEPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the detection method based on PCR technology in the prior art, which has a certain missed detection rate and cannot carry out relative quantification and positioning of the virus in tissue cells.

Method used

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  • Fish probe set for detecting novel coronavirus sars-cov-2 and its preparation method and use
  • Fish probe set for detecting novel coronavirus sars-cov-2 and its preparation method and use
  • Fish probe set for detecting novel coronavirus sars-cov-2 and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The novel coronavirus (SARS-CoV-2) genome (Genebank accession number MN988668.1) with a relatively conserved sequence was selected as the template.

[0055] The present embodiment provides a kind of FISH probe group that detects SARS-CoV-2, comprises:

[0056] 1) The first fluorescent probe group targets the S gene, including DNA probes with nucleotide sequences as described in SEQ ID NO.1-SEQID NO.5; 2) The second fluorescent probe group uses E Gene as the target, including DNA probes with nucleotide sequences as shown in SEQ ID NO.6-SEQ ID NO.9; 3) The third fluorescent probe group, targeting the M gene, including nucleotide sequences as shown in The DNA probe shown in SEQ ID NO.10-SEQ ID NO.13; 4) The fourth fluorescent probe group, with the ORF 3a gene as the target, including nucleotide sequences such as SEQ ID NO.14-SEQ ID NO .17 the DNA probe; 5) the fifth fluorescent probe group, with the N gene as the target, including DNA probes whose nucleotide sequences are...

Embodiment 2

[0068] This embodiment provides a FISH probe set, including: 1) the first fluorescent probe set, targeting the S gene, including RNAs whose nucleotide sequences are respectively described in SEQ ID NO.33-SEQ ID NO.37 Probe; 2) The second fluorescent probe group, which targets the E gene, includes RNA probes whose nucleotide sequences are respectively shown in SEQ ID NO.38-SEQ ID NO.41; 3) The third fluorescent probe The group targets the M gene and includes RNA probes whose nucleotide sequences are shown in SEQ ID NO.42-SEQ ID NO.45; 4) the fourth fluorescent probe group targets the ORF 3a gene and includes The nucleotide sequences are RNA probes shown in SEQ ID NO.46-SEQ ID NO.49; 5) The fifth fluorescent probe group, which targets the N gene, includes nucleotide sequences such as SEQ ID NO. 50- the RNA probe shown in SEQ ID NO.54; 6) the sixth fluorescent probe group, which targets the ORF 1ab gene, including the nucleotide sequences shown in SEQ ID NO.55-SEQ ID NO.62 respec...

Embodiment 3

[0072] This embodiment provides a method for preparing the FISH probe set in Example 1, comprising the following steps:

[0073] S1, designing the probe sequence of the first fluorescent probe group complementary to the S gene sequence, the probe sequence of the second fluorescent probe group complementary to the E gene sequence, and the third fluorescent probe group complementary to the M gene sequence Probe sequence, the probe sequence of the fourth fluorescent probe group complementary to the ORF 3a gene sequence, the fifth fluorescent probe group complementary to the N gene sequence, and the sixth fluorescent probe group complementary to the ORF1ab gene sequence A set of at least two fluorescent probes.

[0074] S2, optimizing the probe sequences of the first fluorescent probe group, the second fluorescent probe group, the third fluorescent probe group, the fourth fluorescent probe group, the fifth fluorescent probe group and the sixth fluorescent probe group Finally, DNA...

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Abstract

The invention discloses a FISH probe group for detecting novel coronavirus SARS-CoV-2, comprising a first fluorescent probe group targeting the S gene, a second fluorescent probe group targeting the E gene, and a fluorescent probe group targeting the M gene. The third fluorescent probe group targeting the gene, the fourth fluorescent probe group targeting the ORF 3a gene, the fifth fluorescent probe group targeting the N gene, and the sixth fluorescent probe group targeting the ORF 1ab gene Needle at least two fluorescent probe groups in the group; in the at least two fluorescent probe groups, the fluorescent molecule labeled by at least one fluorescent probe group has a fluorescence emission spectrum that is different from the rest of the fluorescent probe groups. The FISH probe set has high specificity and sensitivity, can realize the localized detection of SARS-CoV-2 in the sample to be tested, and obtain the distribution and relative quantification of SARS-CoV-2, which is an effective supplement to the detection of new coronavirus nucleic acid .

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection, in particular to a FISH probe set, system, chip, and preparation method and use thereof for detecting the novel coronavirus SARS-CoV-2. Background technique [0002] New coronavirus pneumonia (CoronaVirusDisease2019, COVID-19) referred to as new coronary pneumonia, refers to the pneumonia caused by the 2019 new coronavirus infection (SARS-CoV-2). The clinical manifestations of novel coronavirus pneumonia are mainly fever, fatigue, and dry cough, and upper respiratory symptoms such as nasal congestion and runny nose are rare. About half of the patients developed dyspnea more than a week later, and severe cases rapidly progressed to acute respiratory distress syndrome, septic shock, metabolic acidosis that was difficult to correct, and coagulation dysfunction. Some patients had mild onset symptoms without fever, and mostly recovered after 1 week; most patients had a good prognosis, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6841C12Q1/6811C12N15/11C12R1/93
CPCC12Q1/6811C12Q1/6841C12Q1/701C12Q2563/107
Inventor 董长贵张珮琢王婷飞杜永华胡燕
Owner SHANGHAI GENEPHARMA CO LTD
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