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Joint test strip and preparation method thereof

A technology of test strips and joint inspection tests, applied in the field of joint inspection test strips and its preparation, can solve the problems of low test results, easy hook effect, false negatives, etc.

Inactive Publication Date: 2019-01-18
GUANGZHOU WONDFO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The detection concentration of the first type of analyte is relatively high (generally 0.1-1000mg / L), and the detection range is wide. When the sandwich method is often used for detection, the reaction time is required to be short (about 3-5 minutes), and the Sample dilution is relatively high (above 1:10)
Once the reaction time is too long or the sample dilution ratio is low, there will be an obvious hook effect, resulting in seriously low test results and even false negatives
[0006] The detection concentration of the second type of analyte is relatively low (generally 0.001-1000ng / mL), and the sensitivity for detection of low values ​​is high. When the sandwich method is used for detection, the reaction time must be sufficient (more than 10 minutes) and the sample dilution ratio is relatively low. , in order to meet the requirements for detection sensitivity
However, there is a contradiction between reaction time and dilution ratio in the joint detection of PCT and CRP, which is prone to hook effect, resulting in inaccurate detection results

Method used

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  • Joint test strip and preparation method thereof

Examples

Experimental program
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Embodiment 1

[0066] This embodiment provides a C-reactive protein and procalcitonin combined detection test strip and its preparation method.

[0067] 1. Preparation of joint test strips

[0068] (1) Sample pad preparation

[0069] The diluent is a PBS solution containing 0.3% BSA, 1.2% Tween 20, 6% sucrose, 3% sorbitol, 0.08% sodium azide, 0.03M, pH7.4, where "%" refers to the mass percentage.

[0070] In this embodiment, fluorescent microspheres with a diameter of 200-500 nm are mainly used as the marker substance, and the marker substance will generate emitted light with a Stokes shift of about 50-100 nm after being excited by light.

[0071] Activate the labeling substance by conventional methods (sonication for 30-60 seconds), covalently couple it with an antibody with a protein concentration of 0.2-1.0 mg / mL, react at room temperature for 2 hours, centrifuge and wash twice, each time 8000-15000×g , Centrifuge for 10-15 minutes, and finally dissolve the precipitate with PBS contain...

Embodiment 2

[0115] This embodiment is a modification example of embodiment 1, providing a joint inspection test strip and its preparation method. The changes of this example relative to embodiment 1 include the preparation steps of the joint inspection test strip:

[0116] (1) Sample pad preparation

[0117] The diluent is 0.03M, pH7.4 PBS containing 0.1% BSA, 0.5% Tween 20, 0.5% Tween 80, 2% sucrose, 1% sorbitol, 0.05% sodium azide, where "% " refers to mass percentage.

[0118] The labeled PCT antibody, labeled CRP antibody, and labeled quality control protein (labeled anti-chicken IgY) were respectively adjusted according to the proportions of 5%, 5%, and 0.5% (adjusted according to the actual experimental signal intensity and the trend of the reaction curve), using the above-mentioned After the diluent is diluted, it is evenly sprayed on the sample pad with a gold spraying device, and the spraying volume is 2 μL / cm respectively. After the sprayed sample mat is dried under the condit...

Embodiment 3

[0126] This embodiment is a modification example of embodiment 1, providing a joint inspection test strip and its preparation method. The changes of this example relative to embodiment 1 include the preparation steps of the joint inspection test strip:

[0127] (1) Sample pad preparation

[0128]Diluents include: 0.5% BSA, 2% Tween 20, 1% Tween 80, 10% sucrose, 5% sorbitol, 0.1% sodium azide, constant volume with 0.03M, pH7.4 PBS, where "%" means mass percentage.

[0129] The labeled antibody of PCT, the labeled antibody of CRP, and the labeled quality control protein (labeled anti-mouse IgG) were respectively adjusted according to the ratio of 10%, 10%, and 2% (adjusted according to the actual experimental signal intensity and the trend of the reaction curve). After the diluent is diluted, it is evenly sprayed on the sample pad using a gold spraying device. After the sprayed sample pad is dried under the condition of relative humidity <20% and 37°C, it is sealed and dried a...

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Abstract

The invention provides a joint test strip and a preparation method thereof. The test strip comprises a bottom plate, a sample pad, a coating film and absorbent paper, wherein the sample pad, the coating film and the absorbent paper are sequentially lapped and adhered on the bottom plate; the sample pad is sprayed with a marked first-type analyte antibody, a second-type analyte antibody and a marked quality control protein; the coating film comprises a first-type analyte detection area, a second analyte detection area and a quality control area; the first-type analyte detection area is sprayedwith a protein capable of specifically binding the marked first-type analyte antibody; the second-type analyte detection area is sprayed with another second-type analyte antibody which is located at an epitope different from the epitope of the marked second-type analyte antibody; and the quality control area is sprayed with a protein capable of binding the quality control protein. The joint test strip is capable of realizing the simultaneous detection of two types of analytes under a same high dilution ratio and a long reaction time in the same detection system, and effectively avoiding the hook effect, and is accurate and reliable in detection.

Description

technical field [0001] The invention relates to the field of biological detection, and relates to a combined detection test strip capable of simultaneously detecting two substances and a preparation method thereof. Background technique [0002] Immunoassay is currently the most widely used method in biological detection and clinical medical detection methods. It has the characteristics of high accuracy, sensitivity, and specificity, and can be used in many aspects such as disease diagnosis, physiological activity research (hormones, vitamins), pathological research, and microbial identification. In vitro, antibodies can also specifically recognize the corresponding antigen, and after binding to it, there will be a reaction phenomenon that can be detected by naked eyes or with the aid of instruments. This property is the basis of immunoassay methods. The in vitro antigen-antibody reaction has the characteristics of proportionality and stages, that is, the ratio of antigen-a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/68
CPCG01N33/558G01N33/68G01N2333/47G01N2333/4737
Inventor 黄春荣李思丽王继华
Owner GUANGZHOU WONDFO BIOTECH
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