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Method for simultaneously detecting contents of alpha-lactalbumin, beta-lactoglobulin and lactoferrin based on aptamer probe

A technology of lactoglobulin and lactalbumin, which is applied in the field of immunoanalysis of food analysis, can solve the problems of low cost and inability to detect α-lactalbumin at the same time, and achieve the effect of simple operation, short detection time and high sensitivity

Inactive Publication Date: 2019-03-22
NANJING XIANGZHONG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a method for simultaneously detecting the content of α-lactalbumin, β-lactoglobulin and lactoferrin in food based on an aptamer probe, in order to make up for the inability of the existing methods to simultaneously detect α-lactobumin Blank for protein, β-lactoglobulin and lactoferrin, meeting the requirements of rapid, high sensitivity and low cost detection

Method used

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  • Method for simultaneously detecting contents of alpha-lactalbumin, beta-lactoglobulin and lactoferrin based on aptamer probe
  • Method for simultaneously detecting contents of alpha-lactalbumin, beta-lactoglobulin and lactoferrin based on aptamer probe

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Effect test

Embodiment 1

[0039] In this example, the content of α-lactalbumin, β-lactoglobulin and lactoferrin in milk is detected, and the preparation method of the milk sample solution is as follows: take 50 μL ~ 1mL of the milk to be tested in a 10mL EP tube, and use buffer solution Dilute 200 times to obtain the sample solution to be tested; wherein the buffer is PBS buffer.

[0040] A method for simultaneously detecting the content of α-lactalbumin, β-lactoglobulin and lactoferrin in the above-mentioned milk sample solution based on an aptamer probe, comprising the steps of:

[0041] (1) Apply α-lactalbumin, β-lactoglobulin and lactoferrin antibodies in a 96-well plate with a biochip spotter, and fix the 96-well plate at 37°C for 2 hours; Add 100 μL of 10 mg / mL bovine serum albumin buffer solution to each reaction well after fixation to block the chip for 1 h, then wash with PBST 3 times, 5 min each time;

[0042] (2) Add 25 μL of mixed solutions of α-lactalbumin, β-lactoglobulin and lactoferrin s...

Embodiment 2

[0058] A method for simultaneously detecting the content of α-lactalbumin, β-lactoglobulin and lactoferrin in the above-mentioned milk sample solution based on an aptamer probe, comprising the steps of:

[0059] (1) Apply α-lactalbumin, β-lactoglobulin and lactoferrin antibodies in a 96-well plate with a biochip spotter, and fix the 96-well plate at 37°C for 2 hours; Add 100 μL of 10 mg / mL bovine serum albumin buffer solution to each reaction well after fixation to block the chip for 1 h, then wash with PBST 3 times, 5 min each time;

[0060] (2) Add 25 μL of mixed solutions of α-lactalbumin, β-lactoglobulin and lactoferrin standards with different concentration gradients to some of the reaction wells treated in step (1), and add 25 μL of milk samples to the remaining reaction wells solution, and then add 25 μL of aptamer-modified fluorescent microspheres corresponding to α-lactalbumin, β-lactoglobulin, and lactoferrin to each well, react at 25°C for 0.5 h in all reaction well...

Embodiment 3

[0063] A method for simultaneously detecting the content of α-lactalbumin, β-lactoglobulin and lactoferrin in the above-mentioned milk sample solution based on an aptamer probe, comprising the steps of:

[0064] (1) Apply α-lactalbumin, β-lactoglobulin and lactoferrin antibodies in a 96-well plate with a biochip spotter, and fix the 96-well plate at 37°C for 2 hours; Add 100 μL of 10 mg / mL bovine serum albumin buffer solution to each reaction well after fixation to block the chip for 1 h, then wash with PBST 3 times, 5 min each time;

[0065] (2) Add 25 μL of mixed solutions of α-lactalbumin, β-lactoglobulin and lactoferrin standards with different concentration gradients to some of the reaction wells treated in step (1), and add 25 μL of milk samples to the remaining reaction wells solution, and then add 25 μL of aptamer-modified HRP enzymes corresponding to α-lactalbumin, β-lactoglobulin and lactoferrin to each well, react at 25°C for 0.5h in all reaction wells, and then was...

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Abstract

The invention discloses an aptamer probe for detecting alpha-lactalbumin, beta-lactoglobulin and lactoferrin, and belongs to the technical field of IA (immunoassay) of food analysis. The nucleotide sequence of the aptamer probe for detecting alpha-lactalbumin, beta-lactoglobulin and lactoferrin disclosed by the invention is shown as SEQ ID NO. 1-3. According to a method disclosed by the invention,the contents of the alpha-lactalbumin, the beta-lactoglobulin and the lactoferrin can be simultaneously detected quickly and accurately; the method is high in sensitivity, short in detection time andsimple and easy to operate.

Description

technical field [0001] The invention belongs to a variety of protein detection methods, in particular to a method for simultaneously detecting the contents of α-lactalbumin, β-lactoglobulin and lactoferrin based on an aptamer probe, and belongs to the technical field of immune analysis of food analysis. Background technique [0002] Milk and its dairy products are rich in protein and contain almost all the essential amino acids for the human body. In developed countries, dairy protein and its products provide 20%-30% of food protein and are an important source of protein for people. Especially for infants and young children, cow's milk is an important nutrient besides breast milk, but cow's milk is one of the foods that are more likely to cause allergies. Milk allergy has had an increasing impact on the healthy growth of infants and young children. It can cause infantile diarrhea, rash, iron deficiency anemia, stunting, and in severe cases, it can even cause laryngeal edema...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/68
CPCG01N33/558G01N33/6857
Inventor 李周敏李钟卉
Owner NANJING XIANGZHONG BIOTECH
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