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Non-diagnostic detection method for multiple tumor markers based on SERS sensing substrate

A technology of tumor markers and detection methods, which is applied in the field of non-diagnostic detection of various tumor markers, and can solve problems such as complex operations

Active Publication Date: 2021-04-30
BEIJING INFORMATION SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the operation of this method is still relatively complicated, and the process stability still needs to be improved.

Method used

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  • Non-diagnostic detection method for multiple tumor markers based on SERS sensing substrate
  • Non-diagnostic detection method for multiple tumor markers based on SERS sensing substrate

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preparation example Construction

[0027] (1) Preparation steps of nano-gold system

[0028] Add the trisodium citrate solution to the auric acid chloride solution at 85-95°C, keep it warm for 20-30 minutes, cool down, and centrifuge to obtain the supernatant and the nano-gold system; wherein, the supernatant and the nano-gold system The volume ratio is 9-9.3:1.

[0029] In the present invention, the concentration of the chloride gold acid solution is 0.008-0.012wt%, preferably 0.009-0.011wt%. The concentration of the trisodium citrate solution is 0.9-1.2 wt%, preferably 0.9-1.1 wt%. The volume ratio of trisodium citrate solution to auric acid chloride solution is 1:98-102, preferably 1:99-101. This is conducive to the formation of a stable nano-gold system.

[0030] In the present invention, the auric acid chloride solution is first heated to 85-95° C., preferably 90-95° C., and then the trisodium citrate solution is quickly added under stirring.

[0031] In the present invention, the reaction temperature ...

Embodiment 1

[0070] (1) 50mL concentration is that 0.01wt% auric acid chloride solution is heated to 90 ℃; Add 0.5mL concentration rapidly in stirring process and be the trisodium citrate solution of 1wt%, keep warm reaction 25min under 90 ℃, be down to room temperature, Centrifuge, extract the supernatant until the remaining liquid volume is 5mL, and the remaining liquid is the nano-gold system with a particle size of 40-60nm;

[0071] (2) Mix the above 1mL nano-gold system with 10uL concentration of 10 -3 The 4-mercaptobenzoic acid solution of M was mixed and reacted for 5 minutes, and allowed to stand at room temperature for 30 minutes to obtain a nano-gold-label molecular system;

[0072] (3) Mix the protein spotting solution with the tumor marker to be tested at a volume ratio of 1:9, and place the array on a quartz plate with a uniform grid through the spotting instrument, and one grid corresponds to one array (such as figure 1 shown), and the concentrations of antigens in the obtai...

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Abstract

The invention discloses a non-diagnostic detection method of multiple tumor markers based on an SERS sensing substrate. The detection method comprises the following steps of: (1) adding a trisodium citrate solution into an auric chloride acid solution at 85-95 DEG C, carrying out heat preservation reaction, cooling and centrifuging to obtain a nanogold system; (2) mixing and reacting the nanogold system with a solution containing Raman tag molecules while stirring, and standing to obtain a nanogold tag molecule system; (3) mixing a protein sample application liquid with a tumor marker according to a volume ratio of 1: (8.7-9.3), carrying out sample application in a uniform array, and standing to obtain an antigen array chip; (4) adding an antibody corresponding to the tumor marker into the nano gold tag molecular system for mixing reaction to obtain a nanogold- tag molecular-antibody system; (5) sealing a point sample in the antigen array chip; (6) performing antigen-antibody reaction; and (7) performing Raman scanning imaging. The method is simple to operate and good in process stability.

Description

technical field [0001] The invention relates to the technical field of tumor marker detection, in particular to a non-diagnostic detection method for various tumor markers based on a SERS sensing substrate. Background technique [0002] Tumor markers, also known as tumor markers, refer to substances that are characteristically present in malignant tumor cells, or abnormally produced by malignant tumor cells, or produced by the host in response to tumor stimuli, and can reflect the occurrence and development of tumors. A class of substances that monitor tumor response to therapy. Tumor markers exist in the tissues, body fluids and excreta of tumor patients, and can be detected by immunological, biological and chemical methods. Generally, one cancer has multiple related tumor markers, and one tumor marker may express several cancers. In order to avoid erroneous detection caused by differences in expression in different people, the simultaneous detection of multiple tumor mar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/543G01N21/65
CPCG01N33/57484G01N33/54313G01N21/658Y02A50/30
Inventor 程琳朱桂贤刘淦楠许佳琦
Owner BEIJING INFORMATION SCI & TECH UNIV