Qualitative and quantitative detection dual-purpose HCG test paper for colloidal gold immunochromatography assay

A technology for detecting test strips and immunochromatography, which is applied in the field of in vitro immunodiagnostic reagents to achieve the effects of small coefficient of variation, high detection accuracy, and avoiding inconvenience and errors.

Inactive Publication Date: 2010-09-08
沈鹤柏
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the detection test paper in the prior art is difficult to realize qualitative and quantitative detection on the same test paper

Method used

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  • Qualitative and quantitative detection dual-purpose HCG test paper for colloidal gold immunochromatography assay
  • Qualitative and quantitative detection dual-purpose HCG test paper for colloidal gold immunochromatography assay
  • Qualitative and quantitative detection dual-purpose HCG test paper for colloidal gold immunochromatography assay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The preparation of embodiment 1 human chorionic gonadotropin (HCG) immunochromatography test paper

[0049] 1. Preparation of colloidal gold-labeled HCG monoclonal antibody complex

[0050] Label the HCG monoclonal antibody on the surface of the prepared colloidal gold particles with an average particle size of 3nm-80nm, wash with 0.002M boric acid buffer (containing 1.0% sucrose, 1% BSA) and concentrate to 1 / 2 of the original volume -1 / 50, generally 1 / 10, stored at 4°C.

[0051] 2. Preparation of immunochromatographic test paper

[0052] 2.1 Preparation of sample pad: select glass cellulose membrane as the sample pad material, soak it in the sample pad treatment solution (0.01mol / LPBS (pH=7.4), containing 1% BSA, 0.1% Triton X-100), Dry at 30-42°C, generally at 37°C for later use.

[0053] 2.2 Preparation of conjugation pad: glass cellulose membrane was selected as the conjugation pad material, and the colloidal gold-HCG antibody complex prepared above was redispers...

Embodiment 2

[0056] Example 2 Qualitative detection of 60 clinical patient urine samples and result judgment

[0057] Take 50 positive urine samples and 10 negative urine samples as the samples to be tested, drop them into the sample holes of the test card respectively, and let it stand for 10 minutes. As a result, 23 samples appear red at the qualitative line and the quality control line. It is a positive sample; 27 samples all appear red in the three lines of qualitative line, quality control line and quantitative line, which are strong positive samples; 10 samples only appear red in the quality control band, which is a negative sample (HCG concentration is less than 25IU / L). In the test, if the quality control strip does not develop color, it is invalid.

Embodiment 3

[0058] Example 3 Quantitative detection of 25 clinical patient samples

[0059] 1. Use normal human urine as the diluent for the standard solution of HCG antigen, and configure a series of concentration standard products as follows: 0IU / L, 25IU / L, 100IU / L, 300IU / L, 500IU / L, 800IU / L, 1000IU / L, 3000IU / L, 5000IU / L, 8000IU / L, 10000IU / L, and 15000IU / L.

[0060] 2. Add the standard substance to be tested dropwise into the sampling hole of the detection card, and read the ratio of the signal intensity of the quantitative line to the quality control line through a quantitative detector after 10 minutes (each sample is measured 3 times with 3 detection cards, Take the average value) and draw the corresponding standard curve.

[0061] 3. Use the above standard curve to set the built-in standard curve of the immunochromatography quantitative detector.

[0062] 4. Detect 25 clinical patient samples according to the method in the above step 2, and obtain the content of HCG in the clinic...

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Abstract

The invention relates to qualitative and quantitative detection dual-purpose HCG test paper for colloidal gold immunochromatography assay. The test paper comprises a combined pad and an analysis membrane, wherein HCG monoclonal antibody complex labeled by colloidal gold is distributed on the combined pad; and a qualitative line containing HCG monoclonal antibodies or goat-anti-mouse antibodies, a quality control line and a quantitative line are sequentially distributed on the analysis membrane at fixed points according to a chromatographic flow direction. The invention has the advantages that the qualitative detection can be realized, the sensitivity reaches 25IU/L, the quantitative function is obtained, the situation of barbs caused by too high HCG content of the existing detection test paper is overcome, the linearity range can reach 100-15000IU/L through providing the corresponding quantitative detector for detection under the situation that samples to be detected are not diluted, the inconvenience and the error caused by sample pre-dilution can be effectively avoided, the detection precision is high, the accuracy is high and the requirements on clinical examination are satisfied.

Description

technical field [0001] The invention belongs to the field of in vitro immunodiagnostic reagents. Background technique [0002] Human chorionic gonadotropin (HCG) is a glycoprotein hormone secreted by villous syncytiotrophoblasts, which begins to be produced after implantation in the blastocyst stage. HCG can be detected in the urine of pregnant women 7-10 days after egg cell fertilization, and the concentration of HCG will increase rapidly with the increase of pregnancy, and the doubling time is 1.98 days. Urinary HCG rises slowly in ectopic pregnancy. If HCG cannot continue to increase at a rate of 66% per day during the first 3-6 weeks of pregnancy, the possibility of ectopic pregnancy or threatened abortion should be considered. Therefore, the dynamic quantitative detection of urine HCG can be used as an indicator for follow-up and monitoring of ectopic pregnancy, which is of great significance for the diagnosis of early ectopic pregnancy. [0003] At present, ectopic p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/76G01N33/577G01N33/545G01N33/558
Inventor 赵露晶周志平杨秀梅孙红英沈鹤柏
Owner 沈鹤柏
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