Thin-layer fluorescence focusing method for rapidly detecting single-stranded nucleic acid

A technology of single-stranded nucleic acid and focusing method, which is applied in the field of thin-layer fluorescence focusing for rapid detection of single-stranded nucleic acid, and can solve the problems of low sensitivity and specificity

Pending Publication Date: 2020-12-25
HUADU DISTRICT GUANGZHOU CITY PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Serum IgM and IgG concentrations can be detected by serological methods such as colloidal gold immunochromatography and enzyme-linked immunoassay (ELISA). The IgM concentration can reflect the current infection situation, but its sensitivity and specificity are low

Method used

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  • Thin-layer fluorescence focusing method for rapidly detecting single-stranded nucleic acid
  • Thin-layer fluorescence focusing method for rapidly detecting single-stranded nucleic acid
  • Thin-layer fluorescence focusing method for rapidly detecting single-stranded nucleic acid

Examples

Experimental program
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Effect test

Embodiment

[0030] Example: A thin-layer fluorescent focusing method for rapid detection of single-stranded nucleic acids is used for the detection of novel coronaviruses.

[0031] (1) Prepare experimental components and experimental equipment

[0032] 1. The connection of the capture probe to the solid phase carrier

[0033] Capture probe sequence:

[0034] C1: TAAAACAggCAAACTgAgTTg-3'-NH2

[0035] C2: TTATggAATTTAAgggAAATA-3'-NH2

[0036] C3: AATATTCCAggCACCTTTT-3'-NH2

[0037] C4: TCCCTCTAAgAAgATAATT-3'-NH2

[0038] C5: TACCTAgTTgTgTAgATTgT-3'-NH2

[0039] Covalent coupling of capture probes to magnetic particles 2 (taking 10mg magnetic particles as an example):

[0040] 1) Add 10mg of carboxy-labeled magnetic particles (the magnetic particles should be fully mixed before suction) into a clean 4ml glass bottle, and remove the solution. Solution removal method: place it on a magnet, wait for all the magnetic particles to gather near the magnet after 10 seconds, absorb the liquid,...

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Abstract

The invention relates to a thin-layer fluorescence focusing method for rapidly detecting single-stranded nucleic acid. The thin-layer fluorescence focusing method comprises the following steps that: 1, a capture probe is connected with a solid-phase carrier, herein the solid-phase carrier includes but not limited to magnetic particles, a polystyrene surface, a polylysine surface and a carboxyl modified surface, and the connection includes non-specific adsorption and covalent coupling; step 2, the probe is coupled with chromogenic microspheres, herein the chromogenic microspheres include but are not limited to fluorescent microspheres with various colors, colored microspheres and the like; and step 3, a detection reaction and a signal are read in the reaction card. Compared with the prior art, the method has the advantages that RNA or DNA can be detected within 30 minutes, the sensitivity reaches 1000copies per sample, detection can be performed on site without a central laboratory, anda tool can be provided for detecting pathogens causing outburst public health events.

Description

technical field [0001] The invention relates to the technical field of single-stranded nucleic acid detection, in particular to a thin-layer fluorescence focusing method for rapidly detecting single-stranded nucleic acid. Background technique [0002] When responding to sudden infectious disease public health events, timely and rapid pathogen detection is more important to control the outbreak of infectious diseases. Since the beginning of 2020, the outbreak of the novel coronavirus (COVID-19) around the world urgently requires rapid and accurate detection methods. The standard method for the detection of the new coronavirus is to use respiratory secretions or blood samples as samples, and use real-time RT-PCR, constant temperature amplification, or genome sequencing methods for detection. The specificity of real-time RT-PCR test nucleic acid is relatively high, but the sensitivity of clinical manifestations is low, about 30%, and often requires multiple tests to confirm or...

Claims

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Application Information

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IPC IPC(8): C12Q1/6834
CPCC12Q1/6834C12Q2563/143C12Q2563/149C12Q2563/107
Inventor 杜利军朱应竹谢正顺
Owner HUADU DISTRICT GUANGZHOU CITY PEOPLES HOSPITAL
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