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Test strip based on TdT signal amplification technique and preparation method thereof

A technology of signal amplification and test strips, applied in biological testing, instruments, measuring devices, etc., can solve the problems of low sensitivity and difficult to achieve rapid target analysis.

Active Publication Date: 2020-01-03
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The sensitivity of conventional colloidal gold test strips is low, and it is difficult to achieve rapid analysis of target substances. At the same time, with the improvement of the supervision system and the strengthening of detection efforts, the content of target detection substances in food will become less and less. strip sensitivity puts higher demands on

Method used

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  • Test strip based on TdT signal amplification technique and preparation method thereof
  • Test strip based on TdT signal amplification technique and preparation method thereof
  • Test strip based on TdT signal amplification technique and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Preparation and detection of colloidal gold test strips for the detection of enrofloxacin

[0065] (1) Preparation of colloidal gold

[0066] After 99g of water and 1mL of 10g / L chloroauric acid solution were magnetically stirred and heated to boiling in an oil bath, 3.5mL of 10g / L sodium citrate solution was added to react for 20 minutes, and then the heating was stopped. Stir vigorously and then overnight to obtain colloidal gold, colloidal gold The particle size of the gold nanoparticles was 15 nm and was stored at 4°C for later use.

[0067] (2) Assembly of gold nanoparticles and enrofloxacin aptamer 1 (aptamer1)

[0068] Mix 20 μL of sulfhydryl-modified enrofloxacin aptamer 1 (aptamer1, 100 μmol) with 1 mL of colloidal gold (2.3 nM). After overnight at 25°C, add 113 μL of 0.1 M PB (the final concentration of PB is 0.01 M), and shake at room temperature for 30 min. ; Add PB solution (PB concentration of 0.01M) containing sodium chloride (2M) with a total...

Embodiment 2

[0097] Embodiment 2 Preparation and detection of colloidal gold test strips for potassium ion detection

[0098] (1) Preparation of gold sol: the preparation process is the same as that in Example 1.

[0099] (2) Assembly of gold nanoparticles and potassium aptamer 1: the preparation process is the same as that of Example 1, except that the DNA sequence of potassium aptamer 1 (aptamer1) is: 5'-SH-AAAAAAAAAAGGTTGGT-Biotin-3 ', 5' modified sulfhydryl group.

[0100] (3) Amplification of Au-aptamer1: the preparation process is the same as that of Example 1.

[0101] (4) Preparation of dendritic nano-gold composite solution: the preparation process is the same as that in Example 1.

[0102] (5) Preparation of binding release pad: the preparation process is the same as that of Example 1.

[0103] (6) The preparation process of the incubation solution of streptavidin-biotin-labeled potassium aptamer 2 (aptamer2) is the same as that in Example 1, except that the DNA sequence of po...

Embodiment 3

[0108] Example 3 Preparation and detection of colloidal gold test strips for kanamycin detection

[0109] (1) Preparation of gold sol: the preparation process is the same as that in Example 1.

[0110] (2) Assembly of gold nanoparticles and potassium ion aptamer 1: the preparation process is the same as that of Example 1, except that the DNA sequence of kanamycin aptamer 1 (aptamer1) is: 5'-SH-AAAAAAAAAATGGGGGTTGAG-3 '.

[0111] (3) Amplification of Au-aptamer1: the preparation process is the same as that of Example 1.

[0112] (4) Preparation of dendritic nano-gold composite solution: the preparation process is the same as that in Example 1.

[0113] (5) Preparation of binding release pad: the preparation process is the same as that of Example 1.

[0114] (6) The preparation process of the incubation solution of streptavidin-biotin-labeled kanamycin aptamer 2 (aptamer2) is the same as that in Example 1, except that: kanamycin aptamer 2 (aptamer2) The DNA sequence is: 5'-GCT...

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Abstract

The invention belongs to the field of food safety detection, and specifically relates to a test strip based on TdT signal amplification technique and a preparation method thereof, wherein a combined release pad of the test strip is coated with dendritic gold nanocomposites that comprise a nanogold-sandwich aptamer 1-PolyN1 composite and a nanogold-PolyN2 composite; a reaction film coats the limitof detection of the sandwich aptamer 2 and coats the quality control line of a sequence probe complementary with the PolyN2. The preparation method comprises the following steps of soaking and absorbing a buffer solution containing the dendritic gold nanocomposites by the combined release pad, and drying the combined release pad; respectively coating the solution containing the sandwich aptamer 2marking streptavidin-biotin and the solution containing the sequence probe marking streptavidin-biotin and complementary with the PolyN2 on the reaction film by a film scratching instrument; and fixing the sample pad, the combined release pad, the reaction film and the absorption pad on a backing in sequence in a tomography direction. The test strip detects signals by amplification to improve thedetection sensitivity.

Description

technical field [0001] The invention belongs to the field of food safety detection, in particular to a test strip based on TdT signal amplification technology and a preparation method thereof. Background technique [0002] At present, the detection methods for enrofloxacin mainly include microbial method, four-plate method, photobioreceptor method, high performance liquid chromatography, thin-house chromatography, liquid chromatography-mass spectrometry, supercritical fluid extraction, high-efficiency Capillary electrophoresis, surface plasmon resonance, immunoassay, etc. However, the disadvantages of these methods are that the instrument is expensive, the operation is complicated, the pre-processing of the sample is cumbersome and immediacy. Among them, high performance liquid chromatography is the national standard method, and its detection limit can reach μg / kg level. However, the pretreatment of this method is troublesome, requires professional personnel to operate, th...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/94G01N33/543
CPCG01N33/558G01N33/9446G01N33/54306G01N33/54346Y02A50/30
Inventor 颜娟田润朱福琳
Owner SHANGHAI OCEAN UNIV
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