Bisphenol A and 17beta-estradiol detection kit based on upconversion fluorescence aptamer sensor, application and detection method

A technology of aptamer sensor and detection kit, which is applied in the field of food safety detection and immunoassay, can solve the problems of the influence of complex components, time-consuming sample processing process, high price, etc., and achieve high specificity effect

Active Publication Date: 2018-11-13
INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Large-scale instrumental analysis methods require expensive instruments, time-consuming sample handling processes, complex processes, and a large number of samples
The traditional immunoassay detection method, the antibody is difficult to preserve, the price is expensive, and the detection is easily affected by the complex components in the food, so the method still has certain defects in the detection sensitivity and accuracy

Method used

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  • Bisphenol A and 17beta-estradiol detection kit based on upconversion fluorescence aptamer sensor, application and detection method
  • Bisphenol A and 17beta-estradiol detection kit based on upconversion fluorescence aptamer sensor, application and detection method
  • Bisphenol A and 17beta-estradiol detection kit based on upconversion fluorescence aptamer sensor, application and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] 1. Preparation of aptamers (bisphenol A and estradiol) labeled with upconversion particles (green light and near-infrared light) excited at 808 nm

[0053] 1) Synthesis method of up-conversion particles (green and near-infrared light) excited by 808nm

[0054] ①Wash all glassware with aqua regia, rinse thoroughly with deionized water, and then dry in a 100°C drying oven.

[0055] ② Preparation of rare earth element trifluoroacetic acid precursor: Accurately weigh 10 mmol each of rare earth element oxides (yttrium oxide, ytterbium oxide, erbium oxide, thulium oxide, and neodymium oxide) into three 20mL two-necked bottles, and install a condensation reflux device. Fix it in an oil bath and raise the temperature to 80°C. Add 60mmol trifluoroacetic acid under slow magnetic stirring, and keep the temperature for 24h. After the reaction, dry in a vacuum oven at 60° C. for 12 hours. The obtained rare earth trifluoroacetate is collected and stored in a constant temperature a...

Embodiment 2

[0080] This example is used to illustrate the application method of the up-conversion fluorescent aptasensor for detecting bisphenol A and estradiol.

[0081] 1) Take 500 μL each of aptamer-modified upconverting particles (green light and near-infrared light) to obtain a 0.5 mg / mL mixed particle dispersion, add a certain concentration of diluted (0.01, 0.1, 1, 10, 100, 100ng / mL) bisphenol A and estradiol standard, add 500 μL, and incubate at 37° C. for 1 h with slow shaking.

[0082] 2) After the reaction, centrifuge at 11,000 rpm for 5 min to remove unbound small molecules, wash repeatedly with hybridization buffer, and redissolve in 1 mL of hybridization buffer.

[0083] 3) 500 μL each of bisphenol A and estradiol-modified black phosphorus nano-gold composites (0.5 mg / mL) with complementary chains of regular tetrahedral structures were added, and reacted with vigorous shaking for 1 h.

[0084] 4) After the reaction, centrifuge at 11000rpm for 10min, wash with 1mL of hybrid...

Embodiment 3

[0087] This example is used to illustrate the specificity experiment of the up-converting fluorescent aptasensor for detecting bisphenol A and estradiol.

[0088] 1) Take 500 μL each of aptamer-modified upconverting particles (green light and near-infrared light) to obtain a 0.5 mg / mL mixed particle dispersion, and add (50 ng / mL) 500 μL of estradiol analogs (diethylstilbestrol, 17α -estradiol, estrone, estriol) standards and bisphenol A analogues (biquinone, 2,2-bis(4-hydroxyphenyl)butane, tetrabromobisphenol A, 6F- Bisphenol A) standard substance, slowly shake the reaction at 37°C for 1h.

[0089] 2) After the reaction, centrifuge at 11,000 rpm for 5 min to remove unbound small molecules, wash repeatedly with hybridization buffer, and redissolve in 1 mL of hybridization buffer.

[0090] 3) 500 μL each of bisphenol A and estradiol-modified black phosphorus nano-gold composites (0.5 mg / mL) with complementary chains of regular tetrahedral structures were added, and reacted with...

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Abstract

The invention belongs to the technical field of food safety testing immunoassay and relates to a bisphenol A and 17beta-estradiol detection kit based on an upconversion fluorescence aptamer sensor, anapplication and a detection method. The kit comprises (1) first carboxylated core/shell upconversion nanoparticles NaYF4:Yb,Er@NaYF4:Nd, (2) second carboxylated core/shell upconversion nanoparticlesNaYF4:Yb,Tm@NaYF4:Nd, (3) a target aptamer sequence and a regular tetrahedron structure composition sequence designed according to a complementary strand of the aptamer as well as (4) a black phosphorus nanogold composite dispersion, wherein targets comprise bisphenol A and 17beta-estradiol; the regular tetrahedron structure composition sequence comprises a complementary strand sequence of the aptamer sequence, and the at least part of the complementary strand sequence forms the regular tetrahedron structure sequence. With adoption of the kit or the detection method, bisphenol A and/or 17beta-estradiol can be detected with high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of food safety detection and immune analysis, and more specifically relates to a bisphenol A and estradiol detection kit and application based on an up-conversion fluorescent aptamer sensor and an up-conversion fluorescent aptamer sensor Bisphenol A and estradiol detection method. Background technique [0002] Bisphenol A (Bisphenol A), also known as BPA, is an important raw material for the synthesis of epoxy resin (EP), polystyrene resin (PS), polycarbonate (PC) plastics, widely used in food cans, water bottles, Dental sealants, food packaging and inner coatings. Since the 1960s, BPA has been used to make baby bottles, sip cups and coatings on the inside of milk powder cans. However, BPA can interfere with human and animal endocrine systems, reduce immune function, reduce semen quality, and seriously threaten human health. [0003] Estradiol (17β-Estradiol), also known as E 2 , is a naturally occurring...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64B01J13/06
CPCB01J13/06G01N21/643G01N2021/6432G01N2021/6439
Inventor 白家磊王江高志贤彭媛任舒悦李双宁保安
Owner INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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