A human epididymis protein immunoassay kit and its application method

A human epididymis protein and immune detection technology, which is applied in the field of biological materials and detection, can solve the problem of low sensitivity, achieve low detection limit, improve detection effectiveness and low cost

Inactive Publication Date: 2017-03-29
SUZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a kit for highly sensitive detection of human epididymis protein based on a surface-enhanced Raman spectrum and a composite magnetic material with magnetic properties for the detection of existing human epididymis protein and its use method.

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  • A human epididymis protein immunoassay kit and its application method
  • A human epididymis protein immunoassay kit and its application method
  • A human epididymis protein immunoassay kit and its application method

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Embodiment 1

[0026] 1. Preparation of the substrate

[0027] 1. Preparation of immune gold nanoparticles: first prepare gold nanoparticles with a particle size of 35nm, take 1 mL of 0.01 g / mL HAuCl 4(1%) prepared into 100 mL 1.0×10 -4 g / mL HAuCl 4 (0.01%), heated and boiled under vigorous stirring, quickly added 1mL 1.0×10 -2 g / mL sodium citrate aqueous solution, keep boiling for 15 min, then cool to room temperature while stirring. Add 2 μL of 4-MBA labeled molecules, centrifuge to disperse, and then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) and N-hydroxysuccinimide (NHS ) (the volume ratio of EDC and NHS is 2:1) solution activation, then inject HE4 antibody, incubate for 2-3 h, centrifuge again, disperse and add 10 μL 5% (mass volume ratio) BSA to block the remaining active sites.

[0028] 2. Fe 3 o 4 Synthesis of nanoparticles: 0.65 g FeCl 3 ·6H 2 O and 0.2 g of sodium citrate were added to 20 mL of ethylene glycol (EG), and stirred evenly by ul...

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Abstract

The invention discloses a human epididymis protein immune detection kit and its use method. Its active ingredients include labeled immune gold nanoparticle reagent and Fe3O4@Au immune nanoparticle reagent with a volume ratio of 1:1. Surface-enhanced Raman spectroscopy and core-shell magnetic materials. After activation of carboxyl groups on the surface of gold nanoparticles, inject human epididymis protein antibody to obtain labeled immune gold nanoparticle reagent; after amination modification on the surface of Fe3O4, mix with gold nanoparticles, inject human epididymis protein antibody, and obtain a reagent with good magnetic properties and Fe3O4@Au immune nanoparticle reagent with excellent SERS enhanced activity. The kit provided by the invention has mild conditions for use, and portable detection instruments can be used without large-scale equipment; the detection limit of human epididymis protein can reach femtogram levels, and has high sensitivity; the base of the kit has a recyclable characteristics, reducing the cost of testing.

Description

technical field [0001] The invention relates to a human epididymis protein immune detection kit and a use method thereof, belonging to the technical field of biological materials and detection. Background technique [0002] Ovarian cancer is a common malignant tumor in gynecology, and it is also the type of tumor with the highest mortality rate in gynecology. At present, the diagnosis of ovarian cancer is mainly based on two detection methods. One is transvaginal ultrasonography (TUV), an imaging method used to examine a woman's reproductive organs, including the uterus, ovaries, cervix, and vagina. Despite its widespread use, this method cannot accurately detect whether a mass is benign or malignant. In addition, the method requires an experienced clinical technician to interpret the test results. Another routine detection method is to detect the biomarker CA125 (carbohydrate antigen 125), which is regarded as the "gold standard" for ovarian cancer diagnosis. However, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/532G01N33/68
CPCG01N33/532G01N33/57449G01N33/68
Inventor 姚建林葛明房聪维田中群
Owner SUZHOU UNIV
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