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Microfluidic fluorescence immunoassay chip for rapidly and quantitatively detecting sST2 in whole blood

A quantitative detection, fluorescence immunological technology, applied in the field of immunoassay, can solve the problems of poor sensitivity, linearity, repeatability and quantitative accuracy, repeatability, accuracy and linear range, sensitivity and linear range, etc. Wide, high sensitivity, high sensitivity effect

Pending Publication Date: 2020-06-19
BEIJING LEADMAN BIOCHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Colloidal gold immunochromatography, simple to operate, but poor in sensitivity, linearity, repeatability and quantitative accuracy
The sensitivity of fluorescence immunochromatography is higher than that of colloidal gold method, but it is still lower than that of ELISA method and CLIA method, while the repeatability, accuracy and linear range are poor
It also has the characteristics of poor sensitivity and linear range
ELISA method has mature technology, high sensitivity, low detection cost, but poor repeatability and linear range, large batch-to-batch difference, complicated operation, and long detection time
The CLIA method is superior to other methods in terms of sensitivity, repeatability, linear range, and accuracy, but requires large-scale detection instruments, long detection time, and limited use scenarios

Method used

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  • Microfluidic fluorescence immunoassay chip for rapidly and quantitatively detecting sST2 in whole blood
  • Microfluidic fluorescence immunoassay chip for rapidly and quantitatively detecting sST2 in whole blood
  • Microfluidic fluorescence immunoassay chip for rapidly and quantitatively detecting sST2 in whole blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Composition and preparation of a microfluidic fluorescent immune chip for rapid quantitative detection of sST2 In this implementation example, a microfluidic fluorescent immune chip for rapid quantitative detection of sST2 in whole blood includes a central plate 1 and a bottom plate 9, and the central plate 1 Being of optically transparent material, the bottom plate 9 is located adjacent to the underside of the center plate 1 , the center plate and the bottom plate being joined to each other directly and in a fluid-tight manner by laser welding in the area where they overlap each other around this recess.

[0064] A sample inlet 3 for introducing samples into the chip is provided on one side of the surface of the center plate. The sample inlet is closed in a pressure-tight manner, and enters the first sample mixing area sequentially from the sample inlet 3 through the flow channel. 4. The tracer area and the second sample mixing area, the two sample mixing area...

Embodiment 2

[0088] Example 2 Rapid Quantitative Detection of sST2 Microfluidic Fluorescent Immunochip Drawing Standard Curve Determination and Information Storage

[0089] Take the chip out of storage conditions and equilibrate to room temperature before testing;

[0090] Step 1. Prepare sST2 calibrator with diluent, the concentration is: 0, 5, 20, 50, 150, 300ng / mL

[0091] Step 2. Take 50 μL of the calibrator and add it to the sample inlet of the microfluidic chip. Before each sampling, the pipette head needs to be replaced to avoid cross-contamination. After 10 minutes, read the system detection signal through ResponseIQ. The concentration was detected twice, and the signal value measurement results of the calibrator solution series are shown in Table 1. The regression curve of the calibrator dose-signal value was obtained by four-parameter logic fitting, as shown in Table 1. image 3 shown.

[0092] Table 1

[0093]

Embodiment 3

[0094] Example 3 Methodological Verification of Microfluidic Fluorescent Immunochip for Rapid Quantitative Detection of sST2

[0095] The chip in embodiment 1 is verified according to the conventional manufacturing and verification procedures in the art, and the results are as follows:

[0096] 1. Chip precision measurement

[0097] 1.1 Intra-batch precision analysis

[0098] A batch of chips in Example 1 was used to measure high and low concentration quality control solution series respectively, and 10 chips were measured in parallel, and the intra-assay coefficients of variation were respectively 3.21% and 2.09%. The results are shown in Table 2.

[0099] Table 2

[0100] Target value (ng / mL) Measurement times Intra-analytical CV(%) 30 10 3.21 150 10 2.09

[0101] 1.2 Batch-to-batch precision analysis

[0102] The chips in Example 1 are taken in three batches, and each batch of chips is measured for high and low concentration quality control...

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Abstract

The invention discloses a microfluidic fluorescence immunoassay chip for rapidly and quantitatively detecting sST2 in whole blood, and belongs to the field of immunoassay. The chip comprises a tracingreagent which comprises a cyanine dye Cy5 labeled sST2 monoclonal antibody and a quality control material, wherein the capture reagent contains an sST2 monoclonal antibody and a quality control material monoclonal antibody. According to the chip, an sST2 antigen in blood is labeled by a tracing antibody to form an immune complex, and the immune complex is captured by a capture antibody and emitslight under excitation of exciting light. The chip has high sensitivity and specificity and a wide detection range, and can be used for evaluating the sST2 level of a patient and prompting the heart failure state.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a microfluidic fluorescent immune chip for rapid quantitative detection of sST2 in whole blood. Background technique [0002] Growth-stimulatory expressed gene 2 protein (ST2) is a member of the interleukin 1 (IL-1) receptor family. There are 2 types of ST2 protein, the first is the soluble form (called soluble ST2 or sST2) and the other is the form bound to the cell membrane called ST2 receptor or ST2L. The ligand for ST2 is interleukin-33 (IL-33). Physiologically, the binding of IL-33 to ST2 receptors is a response to myocardial disease or injury, which can down-regulate cardiac function. The cardioprotective signal of IL-33 can be counteracted by the soluble ST2 signal. The mechanism is that soluble ST2 binds to IL-33 and prevents its binding to the ST2 receptor, so it cannot transmit the cardioprotective signal. Thus, in the presence of high concentrations of soluble ST2, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N21/64
CPCG01N33/6893G01N33/6869G01N33/577G01N21/6428G01N2333/7155G01N2800/324
Inventor 王鹏郭闻轩
Owner BEIJING LEADMAN BIOCHEM
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