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Novel coronavirus kit and method for detecting nucleic acid of novel coronavirus by single closed-tube one-step method

A coronavirus and kit technology, applied in the field of biological sciences, to achieve the effect of improving convenience and shortening detection time

Inactive Publication Date: 2020-05-15
NANJING UNIV
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  • Abstract
  • Description
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Problems solved by technology

The amplification process is a variable temperature process that requires repeated thermal cycles and precision instruments

Method used

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  • Novel coronavirus kit and method for detecting nucleic acid of novel coronavirus by single closed-tube one-step method
  • Novel coronavirus kit and method for detecting nucleic acid of novel coronavirus by single closed-tube one-step method
  • Novel coronavirus kit and method for detecting nucleic acid of novel coronavirus by single closed-tube one-step method

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Embodiment 1

[0025] A separate closed-tube one-step detection kit for novel coronavirus nucleic acid of the present invention, comprising primers, padlock probes and RCA standards, the RCA standards having the nucleotide sequence shown in SEQ ID No.1; The padlock probe has the nucleotide sequence shown in SEQ ID No.2; the primer has the nucleotide sequence shown in SEQ ID No.3.

[0026] The kit includes: primers, padlock probes, RCA standards, T4 DNA ligase, Phi29 DNA polymerase, RCA buffer, dNTP, Hemin and chromogenic solution, which includes TMB and H 2 o 2 .

[0027] A method for detecting novel coronavirus nucleic acid in a single closed-tube one-step method driven by 2019-CoV RNA of the present invention comprises the following steps: (1) a phosphorylated linear padlock probe for isothermal amplification, the probe's Both ends can be specifically recognized and bound by the conserved sequence of 2019-CoV RNA (ie RCA standard);

[0028] (2) When the target 2019-CoV RNA exists in the...

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Abstract

The invention discloses a novel coronavirus kit and a method for detecting nucleic acid of the novel coronavirus by a single closed-tube one-step method. The kit for detecting the nucleic acid of thenovel coronavirus by the single closed-tube one-step method comprises a primer, a padlock probe and a PCA standard substance, wherein the PCA standard substance has a nucleotide sequence shown as SEQID No.1; the padlock probe has a nucleotide sequence shown as SEQ ID No.2; and the primer has a nucleotide sequence shown as SEQ ID No.3. According to a field rapid detection product for nucleic acidof 2019-CoV, the detection time is shortened; the convenience degree is improved; the sooner and focused diagnosis is promoted; and rapid diagnosis of suspected patients and field screening of close contacts are realized.

Description

technical field [0001] The invention relates to the technical field of biological sciences, in particular to a kit for novel coronavirus and a method for detecting nucleic acid of novel coronavirus in a single closed-tube one-step method. Background technique [0002] Since the outbreak of the novel coronavirus pneumonia (NCP), early detection and identification of the virus has always been the source of epidemic prevention and control. Value. [0003] The rapid listing and clinical application of real-time quantitative fluorescent PCR (qPCR) nucleic acid detection kits currently in clinical use have played an important role in the prevention and control of NCP epidemics. However, it cannot meet the current rapidly growing demand for screening and diagnosis of a large number of suspected patients and asymptomatic infections. On the one hand, the reagents and equipment required for qPCR are not always easy to obtain, and have special requirements for personnel and places. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844
CPCC12Q1/701C12Q1/6844C12Q2547/101C12Q2531/119C12Q2531/125Y02A50/30
Inventor 项阳焦瑾段成杰刘云飞李根喜
Owner NANJING UNIV
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