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Primer probe composition, kit and method for detecting coronavirus 2019-nCoV

A 2019-ncov, coronavirus technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial assay/inspection, etc., can solve problems such as poor sensitivity, qualitative judgment cannot be absolutely quantitative, etc., and achieve time-saving, high-efficiency and sensitivity. Effect

Inactive Publication Date: 2020-05-01
苏州行知康众生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of the new coronavirus detection kits launched on the market at this stage are detection kits based on fluorescent quantitative qPCR technology. The fastest detection result output time is 4 hours, and only qualitative judgment cannot be absolutely quantitative, and the sensitivity is poor

Method used

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  • Primer probe composition, kit and method for detecting coronavirus 2019-nCoV
  • Primer probe composition, kit and method for detecting coronavirus 2019-nCoV
  • Primer probe composition, kit and method for detecting coronavirus 2019-nCoV

Examples

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Embodiment 1

[0046] The preparation of embodiment one novel coronavirus 2019-nCoV nucleic acid detection kit components

[0047] According to the gene sequence of the new coronavirus 2019-nCoV (as shown in SEQID NO.7) published by the China National Center for Disease Control and Prevention, design target primers and specific probes, and the designed primers and probes can be artificially synthesized according to existing methods .

[0048] Specifically, the primers and probes are as follows:

[0049]

[0050] In this embodiment, the primer probe composition also includes a fluorescent gene and a fluorescent quencher gene. Specifically, the fluorescent gene is selected from at least one of VIC, FAM, HEX, Cy5, Rox or TET; the fluorescent quenching gene is selected from BHQ-1, BHQ-2, BHQ-3, BBQ or TAMRA at least one of . And, the 5' end of the probe is marked with the fluorescent gene, and the 3' end of the probe is marked with the fluorescence quenching gene.

[0051] Admittedly, in ...

Embodiment 2

[0113] Example 2 Detection of Novel Coronavirus 2019-nCoV Nucleic Acid Using Digital PCR

[0114] In this embodiment, the DNA of the sample to be detected is extracted by using the kit shown in the first embodiment, and the target gene amplification fragment is absolutely quantified by digital PCR, which includes the following steps:

[0115] 1) Sample processing

[0116] Types of samples: collect upper respiratory tract specimens (including but not limited to throat swabs, nasal swabs, nasopharyngeal extracts, deep throat sputum); lower respiratory tract specimens: (including but not limited to respiratory tract extracts, bronchial lavage fluid, alveolar lavage fluid, lung tissue biopsy specimens); tissue culture, etc.

[0117] Sample storage conditions: The collected samples should be sent for inspection in time, and those tested within 24 hours are preferably stored at 4°C, and those tested after more than 24 hours are preferably stored at -70°C, and repeated freezing and ...

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Abstract

The invention relates to a primer probe composition, kit and method for detecting new coronavirus 2019-nCoV. A composition one and a composition two are designed as the primer probe composition, a specific primer probe is designed for multiple target genes of the virus based on the digital PCR technology, the digital PCR detection can directly count a number of positive micro-reaction systems of the PCR terminal point, Poisson statistics is used to directly calculate an absolute copy number of a target molecule, the detection of a single target molecule can be realized, the absolute count andquantitative detection of the virus content is achieved, and in addition, the entire detection time does not exceed 3 h, so that the method is time-saving and efficient, and has high sensitivity.

Description

technical field [0001] The invention relates to a primer probe composition, kit and method for detecting novel coronavirus 2019-nCoV, belonging to the field of in vitro diagnostic detection. Background technique [0002] Novel coronavirus, also known as 2019-nCoV, is a virus that can cause respiratory illness. The virus can cause inflammation and a buildup of mucus and fluid in the airways of the lungs (pneumonia). There are many types of coronaviruses, most of which only infect animals but sometimes mutate and infect humans. [0003] 2019-nCoV can cause respiratory diseases and lead to pneumonia. Symptoms of pneumonia include: fever, cough, difficulty breathing, etc. Routes of infection for the virus include: inhaling droplets from an infected person's cough or sneeze; touching an object contaminated with the virus, such as a table or doorknob, and then touching the mouth, nose or eyes; approaching a virus-carrying animal or eating untreated Cooked or undercooked meat or...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/701C12Q1/6851C12Q2531/113C12Q2563/107Y02A50/30
Inventor 朱琦
Owner 苏州行知康众生物科技有限公司
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