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HCG (human chorionic gonadotropin) colloidal gold immunoassay lateral chromatographic test strip and detection method thereof

A technology of immunological lateral flow chromatography and detection method, which is applied in the field of HCG colloidal gold immunological lateral flow chromatography test strips and its detection, can solve the problems of increasing maintenance cost and increasing the cost of biological raw materials, and achieves low detection cost, The effect of overcoming the HOOK effect and improving accuracy

Active Publication Date: 2015-10-21
SHANGHAI UPPER BIO TECH PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, drawing three lines on the NC membrane not only increases the cost of biological raw materials, but also the pump is a relatively expensive component for the film-drawing instrument, thus increasing the cost of purchasing a pump and maintaining the pump

Method used

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  • HCG (human chorionic gonadotropin) colloidal gold immunoassay lateral chromatographic test strip and detection method thereof
  • HCG (human chorionic gonadotropin) colloidal gold immunoassay lateral chromatographic test strip and detection method thereof
  • HCG (human chorionic gonadotropin) colloidal gold immunoassay lateral chromatographic test strip and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation of HCG colloidal gold immunological lateral flow chromatography test strips.

[0039](1) Preparation of T-line solution: Dilute HCG monoclonal antibody to 1-2mg / mL with 10mmol / L phosphate buffer;

[0040] (2) Preparation of C-line solution: dilute HCG antigen to 1-2mg / mL with 10mmol / L phosphate buffer;

[0041] The nitrocellulose membrane 3 is first pasted on the plastic bottom plate with adhesive backing, and then the Fusion5 membrane 2 and absorbent paper 4 are pasted on both ends of the nitrocellulose membrane 3 respectively. Draw a line T on the nitrocellulose membrane 3 and close to the Fusion5 membrane, and draw a line C near the end of the absorbent paper 4. The distance between the T line and the C line is 4.5 mm.

[0042] Spray the T-line solution on the T-line, and spray the C-line solution on the C-line. The final concentration of the T-line antibody is 1-2 mg / ml, the final concentration of the C-line antigen is 1-2 mg / ml, and the spray volume o...

Embodiment 2

[0045] HCG colloidal gold immunological lateral flow chromatography test strip detection method.

[0046] (1) Preparation of gold standard solution:

[0047] The HCG monoclonal antibody to be labeled and the blank colloidal gold solution (the average particle size of the blank colloidal gold is 15-60nm), the labeling amount is 0.005-0.02mg HCG monoclonal antibody / ml colloidal gold solution, and the supernatant is removed by centrifugation (10000rpm, 20min ); the precipitate is blocked with 0.2%-0.5% BSA aqueous solution, and the supernatant is removed by centrifugation (10000rpm, 20min); the pH value is 7.2-7.6 with a concentration of 0.01-0.02mol / L phosphate buffer (containing a concentration of 0.05%-0.5% surfactant) for centrifugal washing; 0.01-0.02mol / L phosphate buffer (0.5-2% BSA and 1-10% sucrose) with a pH of 7.2-7.6 was concentrated to 1 / 10-1 / 5 of the original volume, stored at 4°C for later use, is gold storage.

[0048] (2) deployment:

[0049] Use phosphate buf...

Embodiment 3

[0060] Precision testing

[0061] Using the test method in Example 2, test the standard substances with known concentration values ​​of 25, 1000, 10000 and 100000mIU / ml, and test each group of concentration values ​​10 times. The specific test results are shown in Table 2 below.

[0062] Table 2 precision test result table

[0063] Standard Concentration (mIU / ml)

25

1000

10000

100000

sample 1

25

1010

10097

99760

sample 2

24

1033

9162

109880

sample 3

23

874

11253

114620

Sample 4

25

1085

9048

124420

Sample 5

25

890

8961

106990

Sample 6

24

901

9000

111607

Sample 7

28

853

8937

96970

Sample 8

26

964

9606

105460

Sample 9

29

851

9313

121260

sample 10

27

1020

9104

94760

average

26

948

9448

108573

SD

2 ...

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Abstract

The invention belongs to the field of clinical medical diagnosis and particularly relates to an HCG (human chorionic gonadotropin) colloidal gold immunoassay lateral chromatographic test strip and a detection method thereof. The HCG colloidal gold immunoassay lateral chromatographic test strip consists of a nitrocellulose film, a Fusion 5 film and water-sucking paper, wherein a detection line T line and a quality control line C line are arranged on the nitrocellulose film; a T line solution is sprayed onto the detection line T line; a C line solution is sprayed onto the detection line C line. A standard curve line is made through different concentration values and corresponding T / C average value of a standard product, the standard curve line is subjected to interpolation fit by sample strips for three times, the curved line which is relatively good in linearity and completely raises can be obtained, HOOK effect can be overcome, and detection on high-concentration HCG is realized. The detection range is up to 10 to 200,000mIU / ml, a to-be-detected sample is not required to be diluted, the detection process is simple and convenient, and the detection cost is low.

Description

technical field [0001] The invention belongs to the field of clinical medical diagnosis, in particular to an HCG colloidal gold immunological lateral flow chromatography test strip and a detection method thereof. Background technique [0002] Human chorionic gonadotropin (HCG) is a glycoprotein secreted by placental trophoblast cells. It is composed of α and β dimer glycoproteins. It can be produced after implantation in the blastocyst stage and is produced by syncytiotrophoblasts. synthesis. HCG begins to be secreted on the 6th day after fertilization. On the 7th day after fertilization, it can be detected in the serum and urine of pregnant women, and it increases exponentially. The average concentration doubles in 1.5-2 days, and the rate of increase varies from person to person. Its concentration reaches a peak at 7-10 weeks of pregnancy, about 20,000-100,000mIU / ml, and decreases at 13-15 weeks of pregnancy until the end of pregnancy. If HCG rises slowly, or the HCG inc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/76
CPCG01N33/76G01N2333/59
Inventor 于灵芝杨晶李福刚
Owner SHANGHAI UPPER BIO TECH PHARMA
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