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Colloidal gold test strip for detecting carcinoma of urinary bladder and preparation method of colloidal gold test strip

A technology of colloidal gold test paper and bladder cancer, which is applied in the biological field, can solve the problems of the influence of the technical level of the operator, the inability of patients to monitor the condition, and the difficulty of deployment in primary hospitals, so as to achieve intuitive and reliable judgment, facilitate self-monitoring, and reduce invasiveness. Check the effect

Inactive Publication Date: 2017-09-05
BEIHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the clinical detection of APOA1 protein in urine is mainly detected by ELISA method. ELISA kits are produced by different manufacturers and lack standardization; and the steps are cumbersome, and the results are easily affected by the technical level of the operator; operators need to have certain molecular biology skills It is not easy to develop in primary hospitals; and special instruments such as microplate readers must be used, so patients cannot monitor their own conditions at home

Method used

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  • Colloidal gold test strip for detecting carcinoma of urinary bladder and preparation method of colloidal gold test strip
  • Colloidal gold test strip for detecting carcinoma of urinary bladder and preparation method of colloidal gold test strip
  • Colloidal gold test strip for detecting carcinoma of urinary bladder and preparation method of colloidal gold test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] 1. Preparation of colloidal gold test paper card for detection of bladder cancer

[0056] Such as figure 1 with figure 2 As shown in 2, the test paper card includes a bottom plate 1, a sample pad 2, a colloidal gold pad 3, a chromatography pad (nitrocellulose membrane) 4, and a sample suction pad (absorbent paper or absorbent plate) 5. The base plate 1 is a base plate made of PVC material. Sample pad 2 is made of absorbent glass fibers. The colloidal gold pad 3 is made by spraying the gold-labeled antibody complex on the glass fiber membrane, with a width of 1 cm and a spraying concentration of 30 ug / ml. On the chromatography pad (nitrocellulose membrane) 4, 0.5 cm away from the colloidal gold pad, a detection zone 6 (forming a detection line) consisting of anti-APOA1 monoclonal antibody is sprayed, and the spraying concentration is 100 ug / ml. Distance detection area (detection line) 6 is close to one side 1cm of suction pad, is sprayed with the quality control are...

Embodiment 2

[0066] A method for preparing a colloidal gold test paper card for detecting bladder cancer, comprising:

[0067] 1) Preparation of colloidal gold pads

[0068] a. Pretreatment of gold pads

[0069] Cut the gold pad (made of glass fiber film or polyester film) into a size of 1cm*12cm, soak it in gold pad buffer for 10-15 minutes, dry it, and set aside;

[0070] The gold pad buffer: borate buffer + 2% BSA + 3% sucrose + 0.6M NaCl + 0.2% Tween 20;

[0071] Described borate buffer solution: the H of 0.1Mol / L 3 BO 4 +0.0025Mol / L of Na 2 B 4 o 7 10H 2 O, pH=9;

[0072] The 2% BSA is prepared with PBS, and BSA is bovine serum albumin.

[0073] Studies have found that adding sucrose can effectively slow down the retention of gold-labeled antibodies.

[0074] b. Preparation of colloidal gold solution

[0075] The container is siliconized in advance, that is, the container is soaked in 5% dichloromethylsilane in chloroform solution for 10 minutes, rinsed with triple distille...

Embodiment 3

[0088] The difference between the preparation method of the colloidal gold test paper card for detecting bladder cancer and that of Example 2 is that the detection line and the quality control line are replaced by dots, and the specific implementation method is as follows:

[0089] a. Prepare the quality control line antibody (the secondary antibody of goat anti-APOA1 polyclonal antibody) and the detection line antibody (mouse anti-APOA1 monoclonal antibody) at concentrations of 1mg / ml and 100ug / ml, respectively;

[0090] b. Take 0.3ul of the antibody solution with a pipette gun and drop it onto the NC membrane (nitrocellulose membrane)

[0091] c. Leave it to dry, and store it in a dry place at 4°C in the dark.

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Abstract

The invention relates to a colloidal gold test strip for detecting carcinoma of urinary bladder and a preparation method of the colloidal gold test strip. The test strip comprises a bottom plate, and a sample pad, a colloidal gold pad, a chromatographic pad and a sampling pad which are sequentially adhered onto the bottom plate, wherein the colloidal gold pad and the sampling pad are respectively overlapped at two ends of the chromatographic pad; the other end of the colloidal gold pad is inserted into the sample pad; a gold-labeled antibody complex formed by binding colloidal gold and a rabbit-anti APOA1 (Apolipoprotein) polyclonal antibody is coated on the colloidal gold pad; and secondary antibodies of anti-APOA1 monoclonal antibodies and anti-APOA1 polyclonal antibodies are coated on the chromatographic pad. The colloidal gold test strip disclosed by the invention is capable of realizing detection of the carcinoma of urinary bladder, is fast, high in sensitivity, high in specificity and good in stability, is intuitive and reliable in result judgment and is easily mastered by personnel in grass-roots units, a state of the illness is conveniently monitored by a patient, and invasive inspection of the patient is reduced. The colloidal gold test strip provides a reliable basis for diagnosis of the carcinoma of urinary bladder and is expected to become a novel method for diagnosing the carcinoma of urinary bladder.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a colloidal gold test paper card for detecting bladder cancer and a preparation method thereof. Background technique [0002] At present, the diagnosis of bladder cancer, especially the monitoring of recurrence after treatment, is mainly based on urine exfoliated cytology and cystoscopic biopsy, but urine exfoliated cytology is affected by many factors and the positive rate is low; although cystoscopy is the most reliable diagnostic method , but it is an invasive operation with certain trauma and complications, so it is not easily accepted by patients. APOA1 protein (Apolipoprotein1) is a new bladder cancer marker discovered in recent years. Studies have shown that APOA1 protein is used as a diagnostic indicator for bladder cancer, with a sensitivity of 94.6% and a specificity of 92.0%. At present, the clinical detection of APOA1 protein in urine is mainly detected by ELISA m...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/558
CPCG01N33/558G01N33/57407
Inventor 郑丽沙王又萱呼延楚翘逄冠翔黄重荃宋行文扎西平措郑雨帆樊瑜波
Owner BEIHANG UNIV
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