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Multi-tumor combined detection kit and use method thereof

A combined detection and kit technology, which is applied in the field of multi-tumor joint detection kits, can solve the problems of low detection sensitivity, achieve good repeatability, reduce background interference, and achieve the effects of batch processing

Inactive Publication Date: 2020-06-19
HANGZHOU GUANGKEANDE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The main purpose of this application is to provide a multi-tumor joint detection kit to solve the problems of only one indicator and tumor item at a time and low detection sensitivity in related technologies

Method used

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  • Multi-tumor combined detection kit and use method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 2

[0035]Different types of polymer fluorescent encoded microspheres 7 are coupled with 1 to 100 kinds of second proteins respectively, and then mixed according to a certain ratio to form polymer encoded microsphere suspension A, time-resolved fluorescent microspheres or quantum dot fluorescent microspheres Coupled with streptavidin to form a time-resolved fluorescent microsphere-streptavidin complex. 1 to 100 kinds of first proteins corresponding to macromolecule-encoded microglobulins are respectively cross-linked with biotin to form biotinylated first proteins, and then the biotinylated first proteins are combined with time-resolved fluorescent microglobulins in a certain ratio. The ball-streptavidin complex is mixed to form a time-resolved microsphere suspension B, and then the sample to be tested is added to form a microsphere mixture (polymer fluorescent code-second protein + analyte + streptavidin - Time-resolved fluorescent microspheres (or quantum dot fluorescent microsp...

Embodiment 3

[0038] Using the kit of the present invention to carry out combined detection of multiple tumors, the steps are as follows:

[0039] 1. Polymer fluorescence-encoded microglobulin coupling

[0040] Take 100 microliters of fluorescently encoded microspheres in a 1.5 ml centrifuge tube, centrifuge, wash the pellet with deionized water and activation buffer once (vortex and sonicate for 20 seconds during washing), and resuspend in activation buffer . Add freshly prepared coupling reagents to the microspheres one after another, mix gently, shake slightly at room temperature in the dark, and vortex once in the middle. The microspheres were then centrifuged, the pellet washed with cross-linking buffer, and resuspended in cross-linking buffer. Add protein to the activated microsphere suspension, make up the volume with cross-linking buffer, shake gently at room temperature in the dark, then shake overnight at 4°C. Centrifuge the microspheres, add blocking solution, shake at room te...

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Abstract

The invention discloses a multi-tumor combined detection kit. The multi-tumor combined detection kit comprises fluorescent microspheres capable of performing time resolution, a signal amplification system, a first tumor-associated protein, a sample to-be-detected object, a second tumor-associated protein and polymer fluorescent coding microspheres. The invention further discloses a use method of the kit. The method comprises the following steps: adding a to-be-detected sample; processing incubation oscillation, forming a reaction mixture of macromolecular fluorescence codes, second tumor-associated protein, a to-be-detected object, first tumor-associated protein and fluorescent microspheres capable of being subjected to time resolution; using a time-resolved flow type fluorescence detection device for carrying out high-throughput test; conveying sample liquid and sheath liquid to the flowing chamber together; after to-be-detected liquid particles sequentially pass through a flowing chamber capillary tube detection area, a first laser is activating, enabling a first laser fluorescence detector to collect fluorescence signals while a second laser is activated, and after short-life background fluorescence decay disappears, starting a second laser time-resolved fluorescence detection unit to collect signals for analysis and processing.

Description

technical field [0001] The present application relates to the technical field of biological detection, in particular, to a multi-tumor joint detection kit and its application method. Background technique [0002] The World Health Organization (WHO) and the International Agency for Research on Cancer (IARC) recently released a new report stating that it is estimated that the number of people suffering from cancer in the world is "growing rapidly", with 18.1 million new cases and 18.1 million deaths in 2018 alone 9.6 million. By the end of this century, cancer will become the number one "killer" in the world, and it will also be the biggest "blocker" that hinders human life expectancy and life extension. [0003] Among them, Asia accounted for 48.4% of the world's new cancer cases, equivalent to 1 / 2 of the world's new cases, and Asia accounted for nearly 60% of the 9.6 million cancer deaths. Among the 18.1 million new cancer cases, more than half are men, and there are 9.5 mi...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/543
CPCG01N33/54313G01N33/57484
Inventor 高俊顺高俊莉高金波
Owner HANGZHOU GUANGKEANDE BIOTECHNOLOGY CO LTD
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