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Novel quality control method applied to immunochromatography test paper strip

A quality control method, immunochromatography technology, applied in the direction of analyzing materials, biological testing, material inspection products, etc., can solve the problems of the difference in the coating of the detection line, the influence of the accuracy of the chromatography speed test cannot be ruled out, and simplify the production Effects of processing steps, improving accuracy and precision of measurement, shortening length

Inactive Publication Date: 2017-01-25
GETEIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

3. There are certain differences in the treatment of the bonding pad and the coating of the test line during the processing of the test strips, etc.
Therefore, the test results of the quality control line cannot rule out the influence of the above-mentioned chromatography speed on the test accuracy.

Method used

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  • Novel quality control method applied to immunochromatography test paper strip
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  • Novel quality control method applied to immunochromatography test paper strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation of different wavelength detection molecules and quality control molecule complexes

[0022] 1.1 Preparation of wavelength I detection molecular complex: take 5 mg carboxyl nano fluorescent microsphere I (particle size is 20nm ~ 500nm) according to the conversion of solid content 5mg / ml, add EDC, and its molar content is 1 to 10 times the carboxyl molar content After mixing evenly, shake and react for 30 minutes. After the activation, centrifuge and wash the microspheres with 500 μL of 50 mM PBS solution (pH=7.5). Streptavidin was diluted to 1 mg / mL with 100 mM sodium bicarbonate buffer (pH=8.0), added to the microsphere solution, and placed on a shaker at 220 rpm at 37° C. for 3 h. After the reaction, use 400 μL of 50 mM PBS solution (pH=7.5) to wash the suspended microspheres for three times, and finally add the nanoparticle solution with an amount of 50-500 ug / 100 ul of biotinylated antibody. React for 30-60 minutes, then wash the microspheres ...

Embodiment 2

[0024] Example 2: Preparation of novel immunochromatographic test strips

[0025] Such as figure 2 As shown, a novel immunochromatographic test strip includes a bottom plate 1, on which a sample pad 2, a binding pad 3, a nitrocellulose membrane 4 and a water-absorbing pad 5 are pasted in sequence; wherein the sample pad 2, the binding pad 3 , The nitrocellulose membrane 4 and the water-absorbing pad 5 are overlapped by 1 to 2 mm at the joints to ensure that the test sample passes through the binding pad from the sample area to the detection area smoothly. The preparation method of each part is as follows:

[0026] 2.1 Preparation of sample pad 2: Dissolve 0.5g of BSA protein with 100mM, pH7.2-7.4 PBS buffer solution 100mL, then add 0.01-0.05g of surfactant Tween20 to adjust the pH to 6-8; the sample pad is optional Glass fiber or polyester material, a 30cm long sample pad is placed in 2-5ml of the above-mentioned buffer solution, surfactant and protein mixed solution, soaked...

Embodiment 3

[0032] Example 3: Performance Evaluation of Fluorescence Immunochromatography Diagnostic Reagents

[0033] 3.1 Anti-interference experiment of quality control molecules of different physical examination samples

[0034]Randomly select 80 cases of negative serum and 20 cases of positive serum samples, and use the same sample to drop 100ul / root onto the test strip. For 10 minutes, the fluorescence value of the quality control molecular complex was measured with a Getein 1100 multicolor fluorescent immunoassay analyzer (Nanjing Genetic Biotechnology Co., Ltd.). The measured values ​​of 100 samples, the average value of the sample fluorescence, the average + SD and the average - SD are plotted, and the results are shown in image 3 :

[0035] Such as image 3 As shown, unlike the traditional goat secondary antibody and rabbit secondary antibody which are greatly interfered by different samples, the fluorescence values ​​of the new quality control molecule in different serum qua...

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Abstract

The invention discloses a novel quality control method applied to an immunochromatography test paper strip and belongs to the field of biological detection reagents. The test paper strip comprises a sample pad, a combining pad, a nitrocellulose membrane, water absorbing paper and a backing. The novel quality control method includes: adopting two fluorescent substances different in wavelength to mark respectively detection molecule and quality control molecule I corresponding to analyte to be tested; mixing two fluorescent substance markers according to a certain proportion, and applying to coat the combining pad; applying a mixture of capture molecule and quality control molecule II corresponding to the analyte to be tested to coat an analysis membrane capture line, wherein in the immunoreaction process, the analyte to be tested and the corresponding detection molecule and capture molecule form a sandwiched compound, and the quality control molecule I and the quality control molecule II are specifically combined; calculating a ratio of the compound of the analyte to be tested to a quality control molecule compound. By using the method, interference due to environment factors (temperature, humidity and the like) in the process of chromatography can be eliminated, and the precision and accuracy of measurement can be improved.

Description

technical field [0001] The invention relates to immunochromatography detection technology, in particular to a novel quality control method applied to immunochromatography test strips, and belongs to the field of biological detection reagents. Background technique [0002] Immunochromatography is the use of microporous membrane as a solid-phase carrier, with the mobile phase of the liquid level to be measured, and the tracer markers marked by immunolabeling technology (such as: affinity technology, protein coupling technology, etc.) Lateral flow chromatography is performed by the siphon or capillary action of the membrane. The analyte undergoes a specific immune reaction at the detection line through chromatography, and is detected by naked eyes or corresponding instruments to obtain the judgment result. In recent years, with the increasing demand for basic medical care, home diagnosis and bedside testing, the advantages of immunochromatographic test strips such as simple eq...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/548
CPCG01N33/582G01N33/548
Inventor 黄力杜腾飞朱碧燕苏恩本
Owner GETEIN BIOTECH
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