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Competition method detection test strip based on antigen-aptamer and application thereof

A technology for detecting test strips and aptamers, applied in the field of biomedical detection, can solve the problems of reducing the affinity of complementary chains and aptamers, hybridization of complementary chains, inability to detect, etc., and achieves simple and convenient detection time, simple experimental conditions, and reliable results. Effect

Inactive Publication Date: 2017-01-04
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the aptamer chain is generally relatively long, the affinity between the aptamer and the complementary chain is often greater than the affinity between the aptamer and the target, resulting in the inability to detect; the current literature solution is to truncate the complementary chain, and only partially The complementary chain is immobilized on the detection line, thereby reducing the affinity of the complementary chain and the aptamer, but this method requires complicated exploration, and when the length of the complementary chain is too short, the aptamer cannot hybridize with the complementary chain, which also leads to failure of detection

Method used

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  • Competition method detection test strip based on antigen-aptamer and application thereof
  • Competition method detection test strip based on antigen-aptamer and application thereof
  • Competition method detection test strip based on antigen-aptamer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1 detects the preparation of aflatoxin B1 test strip and its application in detecting aflatoxin B1 (AFB1)

[0063] 1. Preparation of test strips for detecting aflatoxin B1

[0064] 1. Synthesis of detection probes and quality control probes

[0065] Fluorescein-labeled detection probes and biotin-labeled quality control probes were designed and synthesized, respectively.

[0066] The detection probe is the aflatoxin B1 aptamer labeled with fluorescein Cy5 with the connection sequence PolyT18; the aflatoxin B1 aptamer can bind to the aflatoxin B1 hapten in the detection area, and the aflatoxin B1 aptamer The nucleotide sequence of the body is as follows:

[0067] 5'-Cy5-TTT-TTT-TTT-TTT-TTT-TTT-GTT-GGG-CAC-GTG-TTG-TCT-CTC-TGT-GTC-TCG-TGC-CCT-TCG-CTA-GGC-CCA-CA -3'

[0068] The quality control probe is biotin-labeled PolyA18 that can bind to the aflatoxin B1 aptamer junction sequence, and can bind to the detection probe junction sequence PolyT18 through base...

Embodiment 2

[0108] Embodiment 2 detects the preparation of the thrombin test strip and its application in the detection of thrombin

[0109] 1. Preparation of thrombin test strips

[0110] 1. Synthesis of detection probes and quality control probes

[0111] Colloidal gold-labeled detection probes and biotin-labeled quality control probes were designed and synthesized, respectively.

[0112] Wherein the detection probe is colloidal gold-labeled thrombin aptamer with linking sequence PolyT20; the thrombin aptamer can bind to the thrombin antigen in the detection area, and the nucleotide sequence of the thrombin aptamer is as follows:

[0113] 5′-SH-TT TTT TTT TTT TTT TTT TTT TTT GGT TGG TGTGGT TGG-3′

[0114] The colloidal gold synthesis method is: heating and boiling 100mL of HAuCl with a concentration of 1mM 4 solution, under vigorous stirring, 10mL of trisodium citrate solution with a concentration of 38.8mM was added, continued to boil for 20 minutes, then cooled at room temperature,...

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Abstract

The invention relates to the field of biomedicine detection, in particular to a competition method detection test strip based on an antigen-aptamer. The test strip comprises a sample absorption pad, a marker pad, a reaction membrane, a water absorption pad and a base plate. The marker pad is coated with a detection probe. The detection probe is an aptamer of a detected target object marked with an indicator for displaying signal strength. The reaction membrane is provided with a detection area and a quality check area. The detection area is fixedly coated with a target object antigen. The quality check area is fixedly coated with a quality control probe.

Description

technical field [0001] The invention relates to the field of biomedical detection, in particular to an antigen-aptamer-based competition detection test strip and its application. Background technique [0002] Nucleic acid aptamer (Aptamer) is a single-stranded oligonucleotide, mostly single-stranded DNA, with a length between 10 and 100 bp. It is usually passed in vitro from a random sequence oligonucleotide library using a specific target Obtained through screening, as a new type of biorecognition molecule, it can specifically bind to target substances with high affinity. At present, there are hundreds of aptamer targets reported in the literature, and the established aptamer-based detection methods mainly include hundreds of methods such as colorimetry, fluorescence, and electrochemical methods, but so far, aptamer-based There are almost no commercial detection products for the body. [0003] Colloidal gold immunochromatography test strip technology is currently one of t...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543
CPCG01N33/558G01N33/54306
Inventor 陈爱亮朱超张桂兰黄亚飞闫娇白文荟
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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