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Kit for detecting novel-coronavirus-susceptive gene and platform

A technology of susceptibility genes and kits, which is used in the determination/examination of microorganisms, resistance to vector-borne diseases, DNA/RNA fragments, etc.

Active Publication Date: 2020-07-28
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to search for all currently reported potential susceptibility genes of the new coronavirus, and cover all the sites obtained from the summary statistics in the latest databases, guidelines or expert consensus according to the existing research literature and market conditions; Secondly, the results should be accurate and reliable, with high sensitivity and specificity, and the amount of template required should be as small as possible to meet the general screening and detection of the population; in addition, the operation should be simple and fast, the result interpretation should be simple and objective, less error-prone, and the throughput is high and the cost is low. Inexpensive, low requirement on the number of samples, can be checked at any time, suitable for large-scale promotion in different regions of the country

Method used

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  • Kit for detecting novel-coronavirus-susceptive gene and platform
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  • Kit for detecting novel-coronavirus-susceptive gene and platform

Examples

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Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: The primers for detecting virus susceptibility gene mutations were synthesized by Shanghai Bailige Biotechnology Co., Ltd., and the sequences are as follows:

[0042] Amplification primers:

[0043]

[0044]

[0045] Single base extension primers:

[0046]

[0047]

Embodiment 2

[0048] Example 2: The mutant plasmids for detecting virus susceptibility genes were synthesized by Shanghai Sangon Bioengineering Co., Ltd., and there were 5 mutant plasmids (pUC57 clone plasmid+insert sequence), as shown in the following table:

[0049]

[0050]

[0051] Note: *The SNP number, common name and mutation frequency data of the corresponding mutation sites come from the global shared SNP database NCBI dbSNP (https: / / www.ncbi.nlm.nih.gov / snp / ), OMIM database (http: / / www.omim.org / ), the International 1000 Genomes SNP Database (https: / / www.ncbi.nlm.nih.gov / variation / tools / 1000genomes / ), literature, and relevant domestic guidelines and reports in China.

Embodiment 3

[0052] Example 3: Preparation of a mutation detection kit for virus susceptibility genes.

[0053] (1) Time-of-flight mass spectrometry detection system nucleic acid sample pretreatment reagent (Zhejiang Dipu Diagnostic Technology Co., Ltd., article number: 20010100), including the following main components:

[0054]

[0055] (2) Amplification reaction primer premix: a mixture of the nucleotide sequences shown in SEQ ID NO: 1 to 38 in claim 1; the preferred solution is that the primers shown in SEQ ID NO: 1 to 11 The concentration is 0.5 μM, the concentration of primers shown in SEQ ID NO: 12-25 is 1 μM, and the concentration of primers shown in SEQ ID NO: 26-38 is 2 μM;

[0056] (3) Single-base extension reaction primer premix: a mixture of nucleotide sequences shown in SEQ ID NO: 39 to 73 in claim 2, the preferred molar concentration of each extension primer is as follows:

[0057] SEQ ID No.39-42: SEQ ID No.43-46: SEQ ID No.47-49: SEQ ID No.50: SEQ ID No.51-52: SEQ ID N...

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Abstract

The invention discloses a kit for detecting a novel-coronavirus-susceptive gene and a detection platform. The kit comprises primers shown in SEQ ID NO: 1 to SEQ ID NO: 38. The kit disclosed by the invention is high in sensitivity and high in specificity, human gDNA nucleic acid samples (about 5-6 gene copies) to the minimum of 0.01ng / microliter can be detected, and the consumption of samples can be lowered; 2-hole detection can be achieved through 35 loci, and the accuracy reaches 100%; through a repeated-optimized-completed pretreatment reagent, a variety of ingredients can be premixed, the difficulty of system preparing and operating by detection personal during clinical application is simplified greatly, operations are facilitated, and the threshold of use and the difficulty of beginning are effectively lowered; and through being matched with the integrated detection platform, the operation is simple, convenient and rapid, and results are detected and analyzed automatically.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, in particular to a kit for detecting susceptibility genes of novel coronavirus (SARS-CoV-2) based on a matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) platform. Background technique [0002] SARS-CoV-2 is a newly emerged variant of coronavirus, so current clinical research is preliminary. The average incubation period of the virus is about 3-7 days, and the longest does not exceed 14 days. Most patients present mainly with lower respiratory symptoms. Common clinical manifestations include fever, limb weakness, dry cough and other symptoms. Other manifestations include nasal congestion, runny nose, headache, sore throat, hemoptysis, sputum, or diarrhea. Some patients only showed low-grade fever, mild fatigue, etc., without pneumonia. There are also some patients without any clinical manifestations. Severe viral infection can cause a variety o...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6813C12N15/11
CPCC12Q1/6888C12Q1/6813C12Q2600/156C12Q2565/627Y02A50/30
Inventor 张春燕田亚平崔佳奕程昱璇史文杰蒋涛谭旭东桑培培张民杰舒扬王巍郭惠民
Owner GENERAL HOSPITAL OF PLA
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