Human epididymis protein immunoassay kit and using method thereof

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

Inactive Publication Date: 2015-05-06
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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.

Method used

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  • Human epididymis protein immunoassay kit and using method thereof
  • Human epididymis protein immunoassay kit and using method thereof
  • Human epididymis protein immunoassay kit and using method thereof

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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 labeling molecules, and then add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) and N-hydroxysuccinimide ( NHS) (volume ratio of EDC and NHS: 2:1) solution activation, then inject HE4 antibody, incubate for 2-3 h, centrifuge again, disperse and add 10 μL of 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 ultrasonic. 1.2 g of sodium...

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Abstract

The invention discloses a human epididymis protein immunoassay kit and a using method thereof. The effective components of the kit comprise a labelled immune gold nanoparticle reagent and a Fe3O4@Au immune nanoparticle reagent in a volume ratio of 1 to 1 and are materials based on surface enhanced raman spectroscopy and core-shell magnetism. The using method comprises the following steps: after activation treatment on surface carboxyl in the gold nanoparticle, injecting a human epididymis protein antibody to obtain the labelled immune gold nanoparticle reagent; after amination modification on the surface of Fe3O4, mixing Fe3O4 with the gold nanoparticle, and injecting the human epididymis protein antibody to obtain the Fe3O4@Au immune nanoparticle reagent with good magnetism and excellent SERS enhanced activity. The kit is mild in using conditions and is capable of adopting a portable detection instrument without needing large-scale instrument and equipment. According to the kit disclosed by the invention, as the detection limit of human epididymis protein can achieve femto level, the sensitivity is high; as the basement of the kit has the characteristic of recycling use, the detection cost is lowered.

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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IPC IPC(8): G01N33/532G01N33/68
CPCG01N33/532G01N33/57449G01N33/68
Inventor 姚建林葛明房聪维
Owner SUZHOU UNIV
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