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Method and test kit for detecting novel coronavirus

A coronavirus and kit technology, applied in the field of biomedicine, can solve problems such as false negatives, and achieve the effect of getting rid of dependence and achieving absolute quantification.

Pending Publication Date: 2020-07-03
大连晶泰医学检验实验室有限公司 +1
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Problems solved by technology

However, currently limited by the type of testing samples, fluorescent quantitative PCR has a certain proportion of false negatives in the nucleic acid detection of throat swab samples, and the actual detection has higher requirements for the sensitivity and specificity of pathogen detection.

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  • Method and test kit for detecting novel coronavirus
  • Method and test kit for detecting novel coronavirus
  • Method and test kit for detecting novel coronavirus

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Embodiment 1

[0032] The primer and probe combination design for the new coronavirus digital PCR detection system and the establishment of the digital PCR detection system and method.

[0033] 1.1 Primer probe design:

[0034] According to the sequence of human novel coronavirus and the sequence of β-actin gene, the following primers and probe combinations (SEQ ID NO.1-3; SEQ ID NO.4-6) for the detection system of novel coronavirus digital PCR were designed:

[0035] Novel coronavirus upstream primers:

[0036] 1AB-F: 5'-CAAGGTAAACCTTTGGAATTTG-3' (SEQ ID NO.1)

[0037] Downstream primers for 2019-nCoV:

[0038] 1AB-R: 5'-TTGTCCTCACTGCCGT-3' (SEQ ID NO.2)

[0039] Novel coronavirus probe: 1AB-P:5'-FAM-TGCCACTTCTGCTGCTCTTCAACC-BHQ1-3'(SEQ ID NO.3)

[0040] Internal reference gene β-actin upstream primer:

[0041] ACTB-F: 5'-GTCTTCCCCTCCATCGTG-3' (SEQ ID NO.4)

[0042] Downstream primers of internal reference gene β-actin:

[0043] ACTB-R: 5'-GTACTTCAGGGTGAGGATGC-3' (SEQ ID NO.5)

[00...

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Abstract

The invention belongs to the technical field of biomedical treatment, and specifically relates to a method and a test kit for detecting novel coronavirus. The test kit comprises a primer pair and a probe, wherein the sequences of the primer pair and the probe are as shown in SEQ ID NO. 1-6. The digital PCR detection test kit provided by the invention can reach the minimum detection sensitivity of50 copies / ml, realizes accurate quantification of trace pathogen nucleic acid, can detect pathogen nucleic acid from a large number of background signals, eliminates the dependence on a standard curve, does not need formula conversion and realizes absolute quantification of pathogen copies.

Description

technical field [0001] The invention belongs to the field of biomedical technology, and in particular relates to a digital PCR detection system primer set for detection of novel coronavirus, and a digital PCR detection kit including the primer set. Background technique [0002] Novel coronavirus (2019-nCoV) is a new type of coronavirus discovered in human body in December 2019 and is highly contagious and transmissible. The clinical manifestations are fever, fatigue and other systemic symptoms, accompanied by dry cough and dyspnea. etc., can rapidly develop into severe pneumonia, respiratory failure, acute respiratory distress syndrome, etc. Coronaviruses are mainly transmitted through direct contact with secretions or via aerosols and droplets. Laboratory testing methods mainly include virus isolation and nucleic acid detection. The development of nucleic acid detection technology for 2019-nCoV is the key to timely discovery of the source of infection, timely and effectiv...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2600/166C12Q2531/113C12Q2521/107
Inventor 李彤邢晓星刘琦王鹏志冯华华刘利成李杰赵金银
Owner 大连晶泰医学检验实验室有限公司
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