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Rapid detection system for novel coronavirus Omicron mutant strain typing

A coronavirus and mutant technology, applied in the field of rapid detection system for the typing of new coronavirus Omicron mutants, can solve the problems of detection reagent failure, false negative results, false negatives, etc., to achieve good sensitivity and specificity, hardware Low requirements and simple operation

Active Publication Date: 2022-02-25
GUANGDONG HECIN SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] On November 26, 2021, the World Health Organization (WHO) listed Omicron (B.1.1.529) as a Variant of concern (VOC), and VOC was the most dangerous Variant strain, suggesting that it may be highly transmissible or highly pathogenic, and may reduce the effectiveness of current drugs and COVID-19 vaccines
[0004] But at present, because this mutation type was discovered recently, there is no relevant approval for the Omicron typing test kit on the market
However, the existing detection reagents cannot detect Omicron types, and may even face Omicron samples, the detection reagents will fail, and thus lead to false negative test results
And this may be a great challenge to the diagnosis of the disease and the effectiveness of the epidemic prevention policy
[0005] None of the existing new crown nucleic acid detection reagents can quickly detect and type Omicron mutant strains, and may even miss the detection of Omicron mutant strain samples and lead to false negative results
At the same time, all the new crown detection reagents are in liquid form at present, and the liquid form is stored, transported and used, which greatly prolongs the detection time of a single sample

Method used

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  • Rapid detection system for novel coronavirus Omicron mutant strain typing
  • Rapid detection system for novel coronavirus Omicron mutant strain typing
  • Rapid detection system for novel coronavirus Omicron mutant strain typing

Examples

Experimental program
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Effect test

Embodiment 1

[0077] Example 1 Design of primer probes for fluorescent PCR detection of Omicron mutant strains

[0078] In this embodiment, in the Dual ARMS PCR design method developed by the applicant, through a large number of bioinformatics analysis and screening of mutation sites that meet the requirements, primers and probes for nucleic acid typing detection of Omicron mutant strains are designed for these mutation sites .

[0079] First of all, Omicron needs to analyze the mutation site of the gene, including specificity and conservation. Download all uploaded VOC delta GK (B.1.617.2), VUM GH / 490R (B.1.640), VOC Alpha GRY (B.1.1.7), VOC beta GH ( B.1.351), VOC gamma GR (P.1), VOC Lambda GR (C.37), VOC Mu GH (B.1.621) and VOC Omicron GRA (B.1.1.529), including BA.1 and BA. 2 All sequences of both branches. Then, the different mutation sites of Omicron were used as targets, and the Blast sequence comparison analysis was performed with all the above sequences, and the occurrence perce...

Embodiment 2

[0112] Example 2 A fluorescent PCR detection system

[0113] The freeze-dried system for PCR detection includes amplification reaction solution (lyophilized powder), positive control and diluent; the amplification reaction solution includes Mg 2+ , DNA polymerase, MMLV reverse transcriptase, freeze-dried protection solution and dNTPs, primers and probes used for nucleic acid detection of novel coronavirus and Omicron mutant strains in Example 1; simultaneously cooperate with HC800 detection instrument, this detection system also Paraffin oil is required. The preparation formula of the lyophilized powder is shown in Table 3 (dosage for 1 person).

[0114] Table 3 Formula of freeze-dried amplification reaction solution for PCR detection

[0115]

[0116] The lyoprotectant solution contains 2% trehalose, 1% mannitol, 0.9% sodium chloride, 0.2% inositol, 0.75mg / mL PEG and 0.2mg / mL Tween 80.

[0117] Put the aliquoted 2019-nCoV Omicron reaction solution directly into the lyop...

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Abstract

The invention discloses a rapid detection system for novel coronavirus Omicron mutant strain typing. Primer probes in the system are designed based on a bidirectional ARMS PCR technology, six mutation sites of a targeted Omicron mutant strain are detected, the detection accuracy of the Omicron mutant strain is improved to nearly 100%, and the lowest detection limit can be as low as 200 copies / mL. The kit has good sensitivity and specificity; and the whole process can be completed within 30 minutes, operation is easy, rapid and sensitive, the requirement for hardware is low, cost is low, and the kit can be prepared through the technology and used for wide popularization. The detection system is made into a freeze-dried state, so that the detection sensitivity is further improved. An effective technical means is provided for rapid detection and screening of the novel coronavirus Omicron mutant strain, and important significance is achieved.

Description

technical field [0001] The invention belongs to the field of virus rapid detection reagents, in particular to a rapid detection system for typing of novel coronavirus Omicron mutant strains. Background technique [0002] On November 26, 2021, the World Health Organization (WHO) listed Omicron (B.1.1.529) as a Variant of concern (VOC), and VOC was the most dangerous Mutant strains, suggesting that it may have high transmissibility or strong pathogenicity, and may reduce the effectiveness of current drugs and the new crown vaccine. This mutant strain has received widespread attention around the world. Therefore, it is urgent to understand the origin and characteristics of Omicron, and clarify the protection of existing drugs and new crown vaccines, so as to provide a theoretical basis for blocking the spread of Omicron and treating COVID-19 caused by this mutant strain. [0003] According to the WHO report, the first known case of Omicron infection can be traced back to a sp...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6858C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6858C12Q2600/156C12Q2531/113C12Q2521/107C12Q2535/137C12Q2563/107
Inventor 李小锋李晨阳高小龙
Owner GUANGDONG HECIN SCI INC
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