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Kit for specific detection of endometrial cancer

An endometrial cancer and kit technology, applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of patient injury, perforation, weak uterine wall, etc., and achieve low cost, high application value, and easy preservation. Effect

Inactive Publication Date: 2016-08-03
李静
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main clinical manifestation of endometrial cancer is irregular vaginal bleeding. Unlike other tumors such as ovarian cancer, the clinical diagnosis mainly relies on diagnostic curettage. However, patients with breast-feeding or invasive endometrial cancer have weak uterine walls, May cause perforation, causing injury to patient

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Obtaining of embodiment 1 CYPA protein

[0019] The CYPA gene shown in Genbank: NP_066953 was expressed by the conventional eukaryotic expression method in the field, and the corresponding target polypeptide protein was obtained.

Embodiment 2

[0020] Screening and preparation of embodiment 2 nucleic acid aptamers

[0021] Design a random nucleic acid library containing about 20 nucleotides at both ends and 1 nucleotides in the middle as follows:

[0022] 5'-ATCGTCACCATGGATACATG(N39)TTGCACAATTGGACAGTTAC-3'; N39 represents 39 random nucleotides.

[0023] The single-stranded DNA library is amplified into double-stranded DNA, and the product is subjected to 2% agarose gel electrophoresis and then gel-cut to recover and purify; the recovered double-stranded DNA is used as a template to transcribe a single-stranded RNA random library in vitro, and the transcript is purified by PAGE . 75μg RNA library was reverse screened with nitrocellulose membrane to remove membrane-bound RNA molecules, then incubated with 2ugCYPA protein at 37°C for 30min, the reaction solution was filtered through nitrocellulose membrane, and the filter membrane was washed; then the filter membrane was cut into pieces and placed in Boil in elution b...

Embodiment 3

[0038] Example 3 The performance measurement of protein binding suitable gametes

[0039] Take 2.0 μg of aptamers, digest them with calf intestinal alkaline phosphatase (CIP) at 37°C for 1 hour, purify and recover the dephosphorylated RNA; label [γ-32P]ATP on the dephosphorylated RNA by T4 polynucleotide kinase end of RNA molecule. 10nmol of radioactively labeled aptamers were incubated with different concentrations (1-200nM) of CYPA at 37°C for 30min, and the reaction solution of each group was filtered through a nitrocellulose membrane, the filter membrane was washed, dried, and detected by a liquid scintillation counter. For the residual radioactivity, the same sample was measured twice in parallel. Calculate the dissociation constant between each aptamer and the target protein. The result is as follows:

[0040] name

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PUM

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Abstract

The invention discloses a kit for specifically detecting an endometrial cancer .Aptamer has good affinity with human Cyclophilin A protein .By means of the aptamer, Cyclophilin A protein in the blood can be trapped, the endometrial cancer is detected through changes of the content of the aptamer, and the aptamer is prepared into the corresponding kit to be used for endometrial cancer screening .The kit has the advantages of being high in sensitivity and low in cost.

Description

technical field [0001] The invention relates to a kit for detecting endometrial cancer and a detection method thereof. Background technique [0002] As one of the three major malignant tumors of the female reproductive tract, endometrial cancer accounts for 20-30% of malignant tumors of the female reproductive tract. With the development of society and the extension of the average life expectancy of human beings, the incidence of endometrial cancer is increasing year by year and accompanied by a younger trend. If endometrial cancer can be diagnosed early and treated reasonably, its prognosis is generally good. The main clinical manifestation of endometrial cancer is irregular vaginal bleeding. Unlike other tumors such as ovarian cancer, the clinical diagnosis mainly relies on diagnostic curettage. However, patients with breast-feeding or invasive endometrial cancer have weak uterine walls. Perforation may result, causing injury to the patient. [0003] Cyclophilin A prote...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/574C12N15/115
CPCG01N33/68C12N15/115C12N2310/16G01N33/57442G01N2333/47
Inventor 李静
Owner 李静
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