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SERS (Surface Enhanced Raman Scattering) immunochromatography test strip for detecting cow milk allergen alpha-lactalbumin and application thereof

A technology of immunochromatographic test paper and lactalbumin, which is applied in the field of food safety detection, can solve the problems of low detection sensitivity and inaccurate quantification, and achieve the effects of high sensitivity, stable fingerprint information, and easy operation

Pending Publication Date: 2022-04-19
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problems of low detection sensitivity and incapable of accurate quantification of traditional colloidal gold test strips, the present invention provides a SERS immunochromatographic test strip for detecting milk allergen α-lactalbumin, which is obtained by combining SERS and immunochromatographic techniques

Method used

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  • SERS (Surface Enhanced Raman Scattering) immunochromatography test strip for detecting cow milk allergen alpha-lactalbumin and application thereof
  • SERS (Surface Enhanced Raman Scattering) immunochromatography test strip for detecting cow milk allergen alpha-lactalbumin and application thereof
  • SERS (Surface Enhanced Raman Scattering) immunochromatography test strip for detecting cow milk allergen alpha-lactalbumin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] (1) Preparation of silver-coated gold nanoparticles

[0086]Add 100mL of 0.01% chloroauric acid solution into a 250mL conical flask, stir and heat until boiling, and control the reaction temperature at 100-110°C; after the solution boils for 5 minutes, quickly add 2mL of 1% trisodium citrate solution, and continue to cook for 10 minutes. Stop the reaction after the solution is relatively transparent, cool to room temperature, make up to 100mL with ultrapure water, add 100mL ultrapure water to the Erlenmeyer flask again, heat to boiling, add 3mL 1% sodium citrate solution, and use a syringe pump to Add 2 mL of AgNO with a concentration of 10 mM dropwise at a rate of mL / min 3 Solution, continue to heat and stir for 60min, stop heating, cool to room temperature, dilute to 200mL with ultrapure water, centrifuge the prepared solution at a speed of 8000rpm for 15min, remove the supernatant, redissolve with ultrapure water, and store in 4 ℃.

[0087] (2) Preparation of SERS ...

Embodiment 2

[0096] The difference between this embodiment and Example 1 is that during the preparation of silver-coated gold nanoparticles, 2 mL of AgNO with a concentration of 10 mM is added dropwise at a rate of 0.2 mL / min with a syringe pump. 3 Solution, the preparation method that is used to prepare SERS probe in embodiment 2, carries out according to the following steps:

[0097] Mix 1 mL of silver-coated gold nanoparticles with 15 μL of 1 mM Raman beacon molecule 4-MBA, adjust the pH value to 7.5, stir magnetically in a glass bottle for 12 h, centrifuge at 6900 g for 15 min, remove the supernatant, and weigh Suspend in water to form a Raman probe, add 20 μg α-lactalbumin rabbit polyclonal antibody, adjust the pH value to 9.0, shake at room temperature for 4 hours, add BSA to block, shake at room temperature for 8 hours, and obtain the SERS probe, centrifuge at 8000g and remove The supernatant was resuspended with PBS buffer containing 1% BSA.

[0098] The conjugation pad treatment ...

Embodiment 3

[0100] The difference between this embodiment and Example 2 is that 10mMAgNO is added dropwise during the preparation of silver-coated gold nanoparticles. 3 The amount of solution is 4mL, the preparation method that is used to prepare SERS probe in embodiment 3, proceeds according to the following steps:

[0101] Mix 1 mL of silver-coated gold nanoparticles with 20 μL of 1 mM Raman beacon molecule 4-MBA, adjust the pH value to 9.0, stir magnetically in a glass bottle for 8 h, centrifuge at 8000 g for 20 min, remove the supernatant, and weigh Suspend in water to form a Raman probe, add 60 μg α-lactalbumin rabbit polyclonal antibody, adjust the pH value to 8.5, shake at room temperature for 4 hours, add BSA blocking solution, shake at room temperature for 10 hours, and obtain the SERS probe, centrifuge at 8000g Remove the supernatant and resuspend with PBS buffer containing 2% BSA.

[0102] The binding pad treatment solution is PBS buffer with 1% BSA and 1% PVP, the amount of t...

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Abstract

The invention discloses an SERS immunochromatography test strip for detecting cow's milk allergen alpha-lactalbumin and application thereof.The test strip sequentially comprises a bottom plate, a sample pad, a combination pad, an NC film and a water absorption pad, a detection line and a control line are arranged on the NC film, a capture antibody alpha-lactalbumin mouse monoclonal antibody is fixed to the detection line, a goat anti-rabbit second antibody is fixed to the control line, and the detection line and the control line are connected with the sample pad. And an SERS (Surface Enhanced Raman Scattering) probe which is formed by coupling and combining silver-coated gold nanoparticles, Raman beacon molecules and a detection antibody alpha-lactalbumin rabbit polyclonal antibody is sprayed on the combination pad. The SERS probe contains Raman beacon molecules and is captured by a capture antibody to react to form a double-antibody sandwich structure when reaching a detection line, the detection line develops color, the redundant SERS probe is combined with a goat anti-rabbit second antibody on a control line along with buffer solution chromatography, the control line develops color, and the SERS probe is detected by the detection line. The detection sensitivity of the cow milk allergen alpha-lactalbumin can be improved by combining a colorimetric method and a Raman method, and the allergy risk can be accurately evaluated.

Description

technical field [0001] The invention relates to a SERS immunochromatographic test strip, in particular to a SERS immunochromatographic test strip for detecting milk allergen α-lactalbumin and an application thereof, belonging to the technical field of food safety detection. Background technique [0002] Milk is the source of intake of vitamins (vitamin A, vitamin B6) and minerals (calcium). It is an essential nutrient for the growth and development of young children. It plays a key role in human bones, hair, skin and teeth. It is the first food that babies eat. one of the foods. However, cow's milk is also one of the first and most common causes of food allergies in early childhood. Allergic diseases caused by allergens have two notable characteristics. One is that the allergy to food begins in infancy and remains unchanged throughout life; Therefore, some people with cow's milk allergies experience allergic reactions to potentially very small amounts of cow's milk. Most ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558G01N33/543G01N21/65
CPCG01N33/6854G01N33/558G01N33/54346G01N21/658
Inventor 魏新林李靖朱永智徐佳彭斓兰魏康王元凤
Owner SHANGHAI JIAO TONG UNIV
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