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Colloidal gold immunofiltration quantitive detection method and reagent kit

A quantitative detection method and detection kit technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of large variation of detection results, time-consuming and laborious, low efficiency, etc., to achieve quality level improvement, cost reduction, and quality requirements Effect

Active Publication Date: 2013-01-16
SHANGHAI UPPER BIO TECH PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the existing technology, the test results vary greatly, and these problems can only be solved by passively screening raw materials and adding pretreatment processes, which is time-consuming, labor-intensive, low in efficiency, and high in cost, and cannot completely solve the problem.

Method used

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  • Colloidal gold immunofiltration quantitive detection method and reagent kit
  • Colloidal gold immunofiltration quantitive detection method and reagent kit
  • Colloidal gold immunofiltration quantitive detection method and reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 prepares the colloidal gold immunofiltration kit that detects CRP

[0028] (1) Preparation of reaction plate

[0029] a. Assemble the absorbent pad, nitrocellulose membrane and plastic shell into a blank reaction plate. There is a round hole in the center of the plastic case.

[0030] b. Coating: spray quality control line 1, detection line, and quality control line 2 on the designated position of the nitrocellulose membrane in the round hole of the plastic shell.

[0031] The detection line is coated with CRP antibody (primary antibody), the concentration is 0.1-2.0mg / ml; the quality control line 1 and the quality control line are coated with CRP standard samples, the concentration is 0.1-2.0mg / ml;

[0032] c. Drying: Dry at 37 degrees for 2 to 24 hours, put it into an aluminum foil bag after cooling, add a bag of desiccant, and seal it.

[0033] (2) Colloidal gold labeling

[0034] a. Label the CRP monoclonal antibody (secondary antibody) to be labele...

Embodiment 2

[0039] The kit obtained in Example 1 was used for detection.

[0040] Take different concentrations of CRP standard sample solutions for testing. The method is to mix 3 μl of different concentrations of CRP standard sample solutions (5, 10, 25, 50, 100, 150 and 200 mg / L) with 200 μl of gold standard solution, Take 120 μl of the mixed solution and add it dropwise on the nitrocellulose membrane in the round hole of the test paper. After it has penetrated completely, add the washing solution and the blocking solution successively.

[0041] Put the kit into the quantitative reading instrument, and read the CCD gray value of the test line, quality control line 1 and quality control line 2 at the same time. Calculate with the following formula:

[0042] R=T / (C1*C2), or R=T / C

[0043] Among them, T, C1 and C2 are the internal readings of the test line, quality control line 1 and quality control line 2 respectively (in the case of only one quality control line, the internal reading ...

Embodiment 3

[0058] Embodiment 3 detects the colloidal gold immunofiltration kit of ALB

[0059] The colloidal gold immunofiltration kit for detecting ALB was prepared by the same method as in Example 1, and the CRP and monoclonal antibody were replaced with ALB protein and corresponding monoclonal antibody, and the number of quality control points or lines was one.

[0060] Detect with the method identical with embodiment 2, the result is as table 3:

[0061] table 3

[0062] ALB concentration

T1

T2

C1

C2

T1 / C1

T2 / C2

10mg / L

58.2

70.4

100.3

130.4

0.58

0.54

30mg / L

169.1

138.8

104.4

103.6

1.62

1.34

50mg / L

330

279.6

115.4

100.9

2.86

2.77

100mg / L

485.8

455.8

98.1

89.5

4.95

5.09

200mg / L

706.8

671.2

82.4

83.2

8.58

8.07

[0063] T1------...

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Abstract

The invention relates to clinical immunodiagnosis reagents and preparation methods thereof, and discloses a colloidal gold immunofiltration quantitive detection reagent kit and a method for detecting antigens and antibodies by colloidal gold immunofiltration quantitive detection. One or two quality control lines or points which contain known to-be-detected substances are added on the colloidal gold immunofiltration quantitive detection reagent kit. In one reaction, change of color developing degree of detection lines (points) is relevant with change of color developing degree of the quality control lines (points) significantly, deviation of the detection lines (points) can be corrected by the quality control lines (points), and thus, variation caused by deviation of raw materials (nitrocellulose membranes, water absorbent pads), sample viscosity, temperature and sample adding quantity and the like can be overcome effectively.

Description

technical field [0001] The invention relates to a clinical immunodiagnostic reagent and a preparation method thereof, in particular to a colloidal gold immunofiltration technique and a quantitative method thereof. Background technique [0002] In 1971, Faulk and Taylor (Immunochem, 8: 1081, 1971) first used immune colloidal gold as a marker for immunoelectron microscopy. Since then, immune colloidal gold technology has been widely used as a new immunological method. Colloidal gold particles of various sizes can be conveniently prepared from chloroauric acid by reduction method. The method introduced by Frens is the most commonly used (Nature (Phys. Sci.) 241:20, 1973), using trisodium citrate as reducing agent The obtained 10-70nm colloidal gold is the most commonly used size range in clinical diagnostic reagents, and the particle size can be accurately determined by the amount of trisodium citrate added. Colloidal gold particles have a strong adsorption capacity for protei...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558
Inventor 龚杰
Owner SHANGHAI UPPER BIO TECH PHARMA
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