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Cervical cancer screening detection kit

A detection kit and cervical cancer technology, applied in the field of cervical cancer screening detection kits, can solve the problems of increased burden on operators, easy generation of air bubbles, time-consuming and labor-intensive water-based mounting tablets, etc., to save time and labor costs, Effect of avoiding air bubbles, improving detection efficiency and detection quality

Inactive Publication Date: 2018-10-30
FUZHOU MAIXIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Water-based mounting agents are prone to air bubbles, have a certain thickness, and have refractive index problems, which affect the observation of staining results to a certain extent and cause uncertainty in the test results. burden, affecting the efficiency of detection

Method used

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  • Cervical cancer screening detection kit
  • Cervical cancer screening detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Immunocytochemical staining of cervical cell slices by the cervical cancer screening and detection kit

[0021] The kit includes: p16 / Ki-67 mixed primary antibody, (mouse anti-p16 monoclonal antibody and rabbit anti-Ki-67 monoclonal antibody); mixed enzyme-labeled secondary antibody (goat anti-mouse secondary antibody linked to alkaline phosphatase and Goat anti-rabbit secondary antibody linked to horseradish peroxidase); DAB staining solution and AP-Red staining solution.

[0022] Wherein the preparation of AP-Red staining solution includes:

[0023] Chromogen preparation: Dissolve 3.65g of new fuchsin in 50ml of 1.9% hydrochloric acid solution by mass to prepare the first mixed solution, place the first mixed solution in an ice-water bath at 0°C, and slowly add 2.07 g of sodium nitrite to obtain the first suspension, the first suspension is subjected to solid-liquid separation, and after removing the solid, the AP-Red chromogen solution is obtained;

[00...

Embodiment 2

[0063] The difference between Example 2 and Example 1 is that the formulation of the AP-Red staining solution is different.

[0064] The preparation of AP-Red staining solution includes the following steps:

[0065] Chromogen preparation: Dissolve 3.5g of new fuchsin in 45ml of 1.5% hydrochloric acid solution by mass to prepare the first mixed solution, place the first mixed solution in an ice-water bath at 0°C, and slowly add 2g of it within 25 minutes Sodium nitrite, obtain the first suspension, carry out solid-liquid separation to the first suspension, obtain AP-Red chromogen solution;

[0066] Substrate preparation: 0.2g of naphthol AS-BI phosphate was added into 40ml of Tris buffer pH8.2 with a concentration of 18% by mass, mixed to obtain a second mixed solution, and the second mixed solution was mixed with 0.4ml of dimethylformaldehyde Base formamide, 0.14ml of Tween-20 mixed to obtain the AP-Red substrate;

[0067] Preparation of staining solution: Mix AP-Red chromog...

Embodiment 3

[0069] The difference between embodiment 3 and embodiment 1 is that the formulation of AP-Red staining solution is different.

[0070] The preparation of AP-Red staining solution includes the following steps:

[0071] Chromogen preparation: Dissolve 3.7g of new fuchsin in 55ml of 2% hydrochloric acid solution by mass to prepare the first mixed solution, place the first mixed solution in an ice-water bath, add 2.2g of sodium nitrite within 35 minutes, The first suspension is obtained, and the first suspension is subjected to solid-liquid separation to obtain an AP-Red chromogen solution;

[0072] Substrate preparation: 0.4g of naphthol AS-BI phosphate was added into 60ml of Tris buffer pH8.6 with a concentration of 25% by mass, mixed to obtain a second mixed solution, and the second mixed solution was mixed with 0.6ml of dimethylformaldehyde Base formamide, 0.16ml of Tween-20 mixed to obtain AP-Red substrate;

[0073] Preparation of staining solution: Mix AP-Red chromogen wit...

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Abstract

The invention provides a cervical cancer screening detection kit. The kit comprises a p16 / Ki-67 hybrid primary antibody, a hybrid enzyme-labeled secondary antibody and a staining solution, wherein thestaining solution comprises an AP-Red staining solution and a DAB staining solution, and the preparation method of the AP-Red staining solution comprises chromogen preparation, substrate preparationand staining solution preparation. A novel cervical cancer screening detection kit is provided in the technical scheme. The AP-Red staining solution of a novel formula is used in the kit, the chromogen in the staining solution possibly has multiple reactive groups, and the groups can react with groups on tissues so as to form covalent bonds, so that cervical cell slices using the kit can directlypass through organic reagents after staining, a water-based sealing agent does not need to be used for sealing the slices, bubbles are avoided, the thickness is controlled, the time and labor cost canbe saved, observation of staining results is prevented from being influenced, and the detection efficiency and detection quality are improved.

Description

technical field [0001] The invention relates to the field of biomedicine, and the feature relates to a cervical cancer screening and detection kit. Background technique [0002] According to incomplete statistics, there are 130,000 new cases in China every year, and 20,000 to 30,000 women may die of cervical cancer. The new cases of cervical cancer in my country account for 28% of the new cases in the world, and the prevention and control of cervical cancer in China has a long way to go. Cytological examination is currently the main screening method for cervical cancer, p16 / Ki-67 cytological double staining is a newly developed cervical cancer screening technology, its high sensitivity and specificity greatly improved the management of cervical cancer screening triage. [0003] AP-Red staining solution is commonly used in immunocytochemistry. In the immune reaction, the alkaline phosphatase carried by the antibody reacts with the AP-Red staining solution, and finally the a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57411
Inventor 杨清海周洪辉王丹胡滨阳
Owner FUZHOU MAIXIN BIOTECH CO LTD
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