Combined parallel detection method for cardiac failure biomarkers and diagnostic reagent kit

A technology of biomarkers and diagnostic kits, applied in the field of in vitro diagnostic detection methods and diagnostic kits, can solve the problems of poor repeatability, insufficient sensitivity, and cumbersome operation of solid-phase biochip technology, and achieve high sensitivity and stability Good, high-throughput results

Active Publication Date: 2010-09-08
MEDI GENETECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these technologies can only detect one marker at a time, and the operation is cumbersome and the sensitivity is poor, which cannot really meet the needs of clinical diagnosis and detection.
However, solid-phase biochip technology has the disadvantages of poor reproducibility, insufficient sensitivity and cumbersome operation.

Method used

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  • Combined parallel detection method for cardiac failure biomarkers and diagnostic reagent kit
  • Combined parallel detection method for cardiac failure biomarkers and diagnostic reagent kit
  • Combined parallel detection method for cardiac failure biomarkers and diagnostic reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Liquid-phase chip combined parallel detection method for three biomarkers of heart failure

[0044] The specific detection method includes the following steps:

[0045] 1. Activation of the required microspheres:

[0046] 1.1 Vortex the microsphere storage solution at full speed for at least 3 minutes to form a uniform microsphere suspension;

[0047] 1.2 Weigh 10 mg of EDC and S-NHS into two centrifuge tubes;

[0048] 1.3 Dissolve in deionized water to make the final concentration 50mg / ml;

[0049] 1.4 Take 1ml of the microsphere suspension and centrifuge at 10000g for 3min, carefully remove the supernatant;

[0050] 1.5 Add 80 μl of activation buffer to resuspend the microspheres;

[0051] 1.6 Add 10 μl of EDC solution (50 mg / ml) and 10 μl of S-NHS solution (50 mg / ml) respectively, mix well, and incubate at room temperature (15-25° C.) in the dark and shake for 20 minutes.

[0052] 2. Coupling of corresponding capture antibodies to activated microspher...

Embodiment 2

[0097] Example 2: Liquid chip joint parallel detection method for 5 biomarkers of heart failure The specific detection method, steps 1-3 are the same as in Example 1.

[0098] 4. Configuration of Antigen Standards

[0099] 5 kinds of heart failure markers ANP, BNP, NT-proBNP, ET-1, ADM were prepared separately according to the concentration of 10000, 2000, 400, 80, 16, 3.2, 0 pg / ml, and the marker mixture was labeled separately For STD6, ​​STD5, STD4, STD3, STD2, STD1, STD0.

[0100] 5. Preparation of microsphere mixture (mixture I) coupled with capture antibody

[0101] Take the microspheres coated with the capture antibodies of five heart failure markers, as follows: ANP capture antibody microspheres 11, BNP capture antibody microspheres 15, NT-proBNP capture antibody microspheres 21, ET-1 capture antibody microspheres Balls 33 and ADM capture antibody microspheres 37 were mixed in equal proportions so that the final concentration of each microsphere was 200 / μl, and stored...

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Abstract

The invention discloses a combined parallel detection method for cardiac failure biomarkers and a diagnostic reagent kit. The combined parallel detection method is characterized in that the method can detect multiple biomarkers in the same sample at one time, namely can detect fluorescent signals of different microspheres by preparing a liquid phase chip, forming a four-connection complex of a biotin-labeled detection antibody, a cardiac failure biomarker, a capture antibody and a microsphere and combining the four-connection complex and streptavidin-phycoerythrin, thus determining the existent and content of various different cardiac failure biomarkers in the sample to be detected. The invention also discloses components for the diagnostic reagent kit. The method and the reagent kit of the invention have the advantages of high sensitivity, high pass property, quick and accurate detection and the like, and can perform qualitative and quantitative detection on the multiple cardiac failure biomarkers at the same time.

Description

technical field [0001] The invention relates to an in vitro diagnostic detection method and a diagnostic kit, in particular to a liquid-phase chip combined parallel detection method and a diagnostic kit for multiple biomarkers of heart failure. Background technique [0002] Cardiovascular disease is one of the main diseases and major causes of death among Chinese urban population and western developed countries, and heart failure (HF) is a serious threat to human life and health. Biomarkers of heart failure (hereinafter referred to as "heart failure markers") can not only accurately diagnose heart failure and evaluate its severity, but also guide clinical treatment and monitoring of efficacy, and judge the prognosis of heart failure. Therefore, the combined parallel detection of various types of heart failure markers has become inevitable, and the high-throughput rapidity, accuracy and stability of multi-index parallel detection are even more crucial. [0003] Natriuretic p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/546G01N33/74G01N33/68G01N21/64
Inventor 邵棠孙黎
Owner MEDI GENETECH
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