Confining liquid applied to immunocytochemical P16 protein staining and preparation method thereof

Abstract

The invention provides confining liquid applied to immunocytochemical P16 protein staining and a preparation method thereof. The confining liquid is prepared from the following components in parts byweight: 40 to 50 parts of skimmed milk powder, 7 to 10 parts of sodium chloride, 0.1 to 0.3 part of potassium chloride, 1.0 to 2.0 parts of disodium hydrogen phosphate, 0.2 to 0.4 part of potassium dihydrogen phosphate and 1000 parts of distilled water, wherein the confining time of the confining liquid is 15 to 40min and the confining temperature is 32 to 35 DEG C. The confining liquid provided by the invention can be used for confining non-specific background staining and does not cause a condition that a developing signal becomes weak due to excessive confining. Compared with relatively common immunohistochemical confining liquid including BSA (Bull Serum Albumin), casein, animal non-immunized serum and the like, the confining liquid provided by the invention has a better confining effect and relatively low cost; meanwhile, the risk of infecting animal viruses does not exist.

Description

technical field [0001] The invention belongs to the technical field of clinical medical pathology, and in particular relates to a blocking solution applied to immunocytochemical P16 protein staining and a preparation method thereof. Background technique [0002] Cervical cancer is one of the most common malignant tumors of the female reproductive system in gynecology. Its incidence rate is second only to breast cancer and ranks second among female malignant tumors. It seriously threatens women's health and quality of life. China is a country with a high incidence of cervical cancer, accounting for about 1 / 3 of the world's incidence and mortality. Cervical cancer is currently the only cancer whose etiology is clear and can be completely cured through effective early diagnosis and treatment. [0003] Currently, liquid-based cytology is widely used in cervical cancer screening. The specificity of liquid-based cytology is about 90%, but the sensitivity is insufficient, about 50...

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Problems solved by technology

[0003] Currently, liquid-based cytology is widely used in cervical cancer screening. The specificity of liquid-based cytology is about 90%, but the sensitivity is insufficient, about 50%, resulting in about half of the patients being missed.

In order to make up for the shortage of liquid-based cytology, HPV gene detection is used as an auxiliary technology in clinical practice. The sensitivity of HPV gene detection is about 90%, but the specificity is less than 10%, which is easy to cause excessive medical treatment and waste of medical resources.

[0005] In the immunocytochemical reaction, the principle of blocking solution is the same as that of immunohistochemistry, but the difference between cell sheet and tissue sheet makes the blocking solution and blocking time commonly used in immunohistochemistry unsuitable for direct use in immunocytochemistry, as disclosed in patent CN102565393A The immunocytochemical staining kit of the company uses BSA and fetal bovine serum to prepare the blocking solution. The blocking effect of the blocking solution is not mentioned in the article, but in fact the blocking solution can easily lead to incomplete non-specific background staining, or blocking Excessive developing signal becomes weak

Patent CN105241726A discloses that the components of the blocking solution are 45-55g of skimmed milk powder, 0.25-0.29g of potassium dihydrogen phosphate, 1.32-1.52g of disodium hydrogen phosphate, 7-9g of sodium chloride, 0.15-0.25g of potassium chloride, and 1L of distilled water , but the sealing time and temperature of the blocking solution are not disclosed in the document, let alone the sealing effect of the blocking solution

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