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Fluorescent microsphere immunochromatographic testing card for testing five indexes of hepatitis b and method for preparing same

A technology of fluorescent microspheres and immunochromatography, applied in the field of medical testing, can solve problems such as poor anti-solution interference and easy leakage of dyes

Inactive Publication Date: 2010-06-09
WUXI ZODOLABS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this fluorescent nanoparticle also has certain defects, such as easy leakage of dyes, poor ability to resist solution interference, etc., which largely limits the scope of its application.

Method used

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  • Fluorescent microsphere immunochromatographic testing card for testing five indexes of hepatitis b and method for preparing same
  • Fluorescent microsphere immunochromatographic testing card for testing five indexes of hepatitis b and method for preparing same
  • Fluorescent microsphere immunochromatographic testing card for testing five indexes of hepatitis b and method for preparing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Preparation and detection method of isothiocyanate fluorescent microsphere immunochromatography test strip A for detecting HBsAg

[0065] (1) Preparation of fluorescein isothiocyanate-silica-shell-core dual-structure fluorescent microspheres:

[0066] Mix 60ml of absolute ethanol, 2.5ml of ammonia water, 4ml of ethyl orthosilicate, 1ml of ultrapure water, and 1mg of fluorescein isothiocyanate, and react in a constant temperature water bath at 80°C for 6 hours, then add Na 2 SiO 3 Closed microspheres, Na 2 SiO 3 The final concentration of the fluorescent microspheres was 3%, and the size of the fluorescent microspheres was controlled by adding hydrochloric acid to adjust the pH value of the solution, and the fluorescent microspheres with the required particle size were synthesized, and the active groups (-COOH) were modified on the surface of the microspheres.

[0067] (2), preparation of fluorescent microsphere-labeled anti-HBs monoclonal antibody S1 (EDC m...

Embodiment 2

[0090] Example 2 Preparation and Detection Method of Tetramethylrhodamine Isothiocyanate Fluorescent Microspheres Immunochromatography Test Strip B for Detecting HBeAg

[0091] (1), preparation of rhodamine tetramethylisothiocyanate fluorescent microspheres

[0092] Mix 10ml of cyclohexane, 12ml of n-hexanol and 5ml of Triton X-100- uniformly in a ratio of 2:1.2:1, add an appropriate amount of water as a dispersed phase, and after ultrasonic emulsification, add 0.5mg of rhodamine tetramethylisothiocyanate, Stir for 20 minutes, add a certain amount of ammonia water until the solution is clear, add 2.5ml of tetraethyl orthosilicate and stir for 2 minutes, add 2ml of aminoethylaminopropyl polydimethylsiloxane (AEAPS), mix well, stir for 24 hours, and then The microparticles obtained by repeated washing and centrifugation are the prepared rhodamine isothiocyanate fluorescent microspheres.

[0093] (2), preparation of anti-HBe monoclonal antibody labeled with fluorescent microsphe...

Embodiment 3

[0107] Example 3 Preparation and detection method of isothiocyanate fluorescent microsphere immunochromatography test strip C for detecting anti-HBs

[0108] (1) The preparation of fluorescein isothiocyanate-silica shell-core dual-structure fluorescent microspheres is as in Example 1.

[0109] (2), preparation of fluorescent microsphere-labeled HBsAg (EDC method):

[0110] Take 1mg of fluorescent microspheres coated with fluorescein isothiocyanate and centrifuge at 1000×g for 10-15min, collect the precipitate, and adjust the concentration of the microspheres to OD with 0.01M borate buffer solution at pH 4.8 450= 0.2. Then add 90μL 50mg / mL p-ethyl-N,N-dimethylpropylcarbodiimide (EDC), 150μL 5mg / mL nitrogen hydroxysuccinimide (NHS), shake and mix well, and incubate at room temperature for 10-30min Afterwards, centrifuge at 1000×g for 5-15min, dissolve the precipitate with 0.01M pH4.8 borate buffer, and adjust the concentration of microspheres to OD 450 0.2 to 1. Add 1 to 20 ...

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Abstract

The invention discloses a fluorescent microsphere immunochromatographic testing card for testing five indexes of hepatitis b and a method for preparing the same. The testing card comprises a hepatitis b surface antigen test paper strip, a hepatitis b e surface antigen test paper strip, a hepatitis b surface antibody test paper strip, a hepatitis b e surface antibody test paper strip, and a hepatitis b core antibody test paper strip. Each test paper strip is formed by overlapping and bonding filter paper, a sample pad, a glass fiber film spray-coated with fluorescent microspheres, a cellulose nitrate film and water absorption paper on a bottom plate by glue in sequence, wherein the cellulose nitrate film is coated with antigens serving as a testing area and anti-rabbit antibodies serving as a quality control area; and during a test, after emitted fluorescent light passes a filter, the emitted spectrum is collected, accumulated and multiplied by the CCD scanning technology and then converted into a numerical signal, the numerical signal is multiplied by a correction factor, and the strength of the corrected fluorescent light is substituted in a standard curve of a fluorescence analyzer, so that the concentrations of the five indexes of hepatitis b of the sample can be automatically worked out. The test of hepatitis b viruses by the testing card has the characteristics of specificity, sensitivity, simpleness and accuracy.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a fluorescent microsphere immunochromatographic detection card for quantitatively detecting five items of hepatitis B and a preparation method thereof. Background technique [0002] As a kind of infectious disease, hepatitis B has become a kind of infectious disease that everyone talks about. Due to my country's poor medical diagnosis conditions, unhygienic eating habits and weak prevention awareness, there are currently more than 120 million hepatitis B virus carriers in my country, accounting for about 10% of the population, and as many as 110,000 people die of liver cancer every year. Therefore, the prevention and treatment of hepatitis B in my country has a long way to go, and the detection of hepatitis B is a key link in the prevention and treatment of hepatitis B. In biological and medical testing, immunoassays are often used, including radioimmunoassays, enzyme-linked imm...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/577G01N33/533G01N33/558G01N33/52C09K11/02C09K11/06
Inventor 熊勇华徐波魏华史爱武陈雪岚赖卫华
Owner WUXI ZODOLABS BIOTECH
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