Kit for specific detection of endometrial carcinoma

一种子宫内膜癌、试剂盒的技术，应用在生物测试、测量装置、材料检验品等方向，能够解决子宫壁薄弱、患者伤害、穿孔等问题，达到易保存、成本低、高应用价值的效果

Inactive Publication Date: 2016-08-10
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 . 75 μg RNA library was reverse-screened through nitrocellulose membrane to remove membrane-bound RNA molecules, and then incubated with 2ug CYPA protein at 37°C for 30 minutes, the reaction solution was filtered through nitrocellulose membrane, and the filter membrane was washed; then the filter membrane was cut into pieces, Place in elution...

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 specific detection of endometrial carcinoma. An aptamer has high affinity to human Cyclophilin A protein. By utilizing the aptamer, the Cyclophilin A protein in blood can be captured, and the endometrial carcinoma is detected through changes in content of the Cyclophilin A protein; after the aptamer is prepared to be the corresponding kit which is about to be used for screening the endometrial carcinoma. The kit has the advantages high sensitivity and low 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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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/573
CPCC12N15/115C12N2310/16G01N33/573G01N33/57442G01N33/68G01N2333/47G01N2333/99
Inventor 李静
Owner 李静
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