Method for detecting brucella by combining surface enhanced Raman scattering with immunochromatography technology

A surface-enhanced Raman and Brucella technology, applied in Raman scattering, measuring devices, analyzing materials, etc., can solve the problems of low success rate, high risk, low sensitivity, etc., and achieve convenient and fast use and sensitivity. High, specific effect

Pending Publication Date: 2021-10-29
GUANGXI UNIV +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] Through the above analysis, the existing problems and defects of the prior art are: conventional methods require harsh conditions for isolation and identification of pathogens, and are laborious, time-consuming, high risk, and low success rate; among them, PBT and SAT have low specificity and low sensitivity ; CFT is cumbersome to operate, requires high experimental conditions and technical level, and is extremely inconvenient for practical application; PCR detection method requires complex equipment and high cost, and is not suitable for grassroots and on-site detection
[0005] The difficulty of solving the above problems and defects is as follows: the difficulty of traditional technology defects is mainly: long time-consuming; high level of operation technology requirements, professional laboratory technicians are required; complex and expensive instruments and equipment are required; rapid on-site detection cannot be carried out

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  • Method for detecting brucella by combining surface enhanced Raman scattering with immunochromatography technology

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Embodiment 1

[0054] Example 1: Prepare materials

[0055] The method that surface-enhanced Raman scattering combined with immunochromatography technology detection Brucella provided by the embodiment of the present invention comprises the following steps:

[0056] S1, Brucella antibody grouping, the Brucella marker antibody group and the Brucella capture antibody group contained in the obtained pair of Brucella antibodies are stored independently for future use;

[0057] S2, to prepare nanomaterials, first select one of the Brucella antibodies in step S1, and use the sodium citrate reduction method to prepare a 20nm AuNPs colloidal gold solution; then add 8mM DTNB to the AuNPs colloidal gold solution, and stir for 3h; The supernatant was discarded by centrifugation, and resuspended to the original volume with deionized water to prepare Au / DTNB NPs; finally, the Au / DTNB NPs were stirred and heated to boil, and 0.5% (w / v) sodium citrate solution was added, followed by dropwise addition of 0...

Embodiment 2

[0078] The method that surface-enhanced Raman scattering combined with immunochromatography technology detection Brucella provided by the embodiment of the present invention comprises the following steps:

[0079] S1, Brucella antibody grouping, the Brucella marker antibody group and the Brucella capture antibody group contained in the obtained pair of Brucella antibodies are stored independently for future use;

[0080] S2, to prepare nanomaterials, first select one of the Brucella antibodies in step S1, and use the sodium citrate reduction method to prepare a 35nm AuNPs colloidal gold solution; then add 12mM DTNB to the AuNPs colloidal gold solution, and stir for 6h; The supernatant was discarded by centrifugation, and resuspended to the original volume with deionized water to prepare Au / DTNB NPs; finally, Au / DTNBNPs were stirred and heated to boiling, and 1.5% (w / v) sodium citrate solution was added, followed by dropwise addition of 1.5 mM silver nitrate solution, keep boil...

Embodiment 3

[0102] The method that surface-enhanced Raman scattering combined with immunochromatography technology detection Brucella provided by the embodiment of the present invention comprises the following steps:

[0103] S1, Brucella antibody grouping, the Brucella marker antibody group and the Brucella capture antibody group contained in the obtained pair of Brucella antibodies are stored independently for future use;

[0104] S2, preparing nanomaterials, first select one of the Brucella antibodies in step S1, and prepare a 25nm AuNPs colloidal gold solution by sodium citrate reduction method; then add 10mM DTNB to the AuNPs colloidal gold solution, and stir for 4 hours; Discard the supernatant by centrifugation, resuspend to the original volume with deionized water, and prepare Au / DTNB NPs; finally take Au / DTNBNPs and stir and heat to boil, add 1% (w / v) sodium citrate solution, and then dropwise add 1mM Keep boiling the silver nitrate solution for 15 minutes to obtain Au / DTNB@Ag NP...

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Abstract

The invention belongs to the technical field of brucella detection, and discloses a method for detecting brucella by combining surface enhanced Raman scattering with immunochromatography. The method for detecting brucella by combining surface enhanced Raman scattering with immunochromatography comprises the following steps: grouping brucella antibodies; preparing a nano material; preparing an SERS labeled detection probe; preparing a Raman immunochromatography test paper strip; and detecting the performance of the test paper strip. According to the method for detecting the brucella by combining the surface enhanced Raman scattering and the immunochromatography technology, the Raman enhancement technology and the immunochromatography technology are combined, the brucella Raman immunochromatography test paper strip is prepared, and the test paper strip is high in sensitivity, high in specificity and convenient and rapid to use, and has great popularization and application values in clinical rapid diagnosis; and the method is easy and convenient to operate and high in sensitivity, detection can be completed within 15 min, and the method has wide application and popularization prospects in early infection detection of brucella.

Description

technical field [0001] The invention belongs to the technical field of Brucella detection, and in particular relates to a method for detecting Brucella by surface-enhanced Raman scattering combined with immunochromatography technology. Background technique [0002] At present, brucellosis is caused by Brucella, which is a widely prevalent and harmful zoonotic disease in the world, causing miscarriage, infertility and local lesions of various tissues. Brucella can be infected through the nose, pharynx, and oral cavity, mainly through the infiltration of mucosal epithelial tissues. At present, there are more than 10 species of Brucella in the genus Brucella. Different species of Brucella have obvious host hazard tendencies, but most of them have the ability of cross-infection between different hosts, which can cause serious diseases. public health issues. With the rapid development of animal husbandry in my country in recent years, brucellosis has become one of the serious d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65G01N33/531G01N33/577
CPCG01N21/658G01N33/531G01N33/577
Inventor 潘艳韦英明陈海兰王冬英蒋钦杨郑自华陈集成
Owner GUANGXI UNIV
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