Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of klk13 in the preparation of a detection kit for diagnosing cervical adenocarcinoma

A detection kit and technology for cervical adenocarcinoma, applied in the field of immune detection, can solve the problem of not finding the specificity of KLK13 expression, and achieve the effect of high sensitivity and specificity, high application prospect, and help in diagnosis

Active Publication Date: 2021-10-19
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
View PDF12 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The specific expression of KLK13 in cervical adenocarcinoma tissue mucus has not been found in this field

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of klk13 in the preparation of a detection kit for diagnosing cervical adenocarcinoma
  • Application of klk13 in the preparation of a detection kit for diagnosing cervical adenocarcinoma
  • Application of klk13 in the preparation of a detection kit for diagnosing cervical adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Effect of Biopsy and Cervical Mucus Samples on EP and PR

[0017] This experiment adopts the EnVision System two-step method: the EnVision chromogenic system connects the secondary antibody and the enzyme into a polymer through a polydextran backbone to directly amplify the signal 40 to 50 times, and the traditional secondary antibody and tertiary antibody are respectively Incubation was combined into one incubation. The cervical tissues of the subjects were fixed with 4% paraformaldehyde fixative solution, dehydrated, transparentized, soaked in wax, and sectioned, followed by two-step immunohistochemical staining, and then used a microscope for diagnosis and image collection. As shown in Table 1, using traditional cervical adenocarcinoma biopsy markers and immunohistochemical detection methods to analyze ER and PR, it can be seen that the specificity and sensitivity of the two for cervical tissue detection are very low. figure 1 The immunohistochemical photo...

Embodiment 2

[0021] Example 2 iTRAQ quantitative analysis test

[0022] 1. Sample Processing and Protein Extraction

[0023] The cervical mucus samples of 9 subjects were extracted by scraping with cotton swabs, among which 3 were normal subjects, 3 were patients with cervical adenocarcinoma in situ, and 3 were patients with invasive cervical adenocarcinoma.

[0024] Add protein lysate (7M Urea / 2M Thiourea / 4% SDS / 40 mM Tris-HCl, pH8.5 / 1mM PMSF / 2mM EDTA) to the sample, mix well and incubate on ice for 5 min, then add the final concentration of 10 mM DTT, sonicate in an ice bath for 15 min, then centrifuge at 13,000 g at 4 °C for 20 min, transfer the supernatant to a new centrifuge tube; Let stand overnight. The protein precipitate was collected by centrifugation and air dried. Add 8 Murea / 100 mM TEAB (pH 8.0) solution to redissolve the protein, add DTT to a final concentration of 10 mM, and carry out the reduction reaction in a water bath at 56 °C for 30 min. Subsequently, IAM was add...

Embodiment 3

[0035] Example 3 siRNA interference cell experiment

[0036] 1. Cell Scratch Test

[0037] The samples in this experiment were cervical tissue mucus from patients with cervical cancer, and the control group was cervical adenocarcinoma HeLa cells whose expression of KLK13 was not knocked down. First, cervical adenocarcinoma HeLa cells were seeded in 96-well plates and cultured in DMEM containing 10% fetal bovine serum (Gibco, USA) at 37°C, 5% CO 2 Routine culture in the incubator, when the degree of cell confluence reached 100%, the mixture of liposome and KLK13 siRNA (5'-UCAAAGUUUACCUAGGCAATT-3', synthesized by Shanghai Gemma Company) was prepared according to the instructions of Lipofectamine 2000 (Invitrogen, USA). HeLa cells were transiently transfected. After 6 h, the transfection reagent was discarded, and DMEM medium containing 10% fetal bovine serum was added to continue culturing for 48 h.

[0038] Scratch the bottom of the 96-well plate with the tip of a plastic ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses the application of KLK13 in the preparation of a detection kit for diagnosing cervical adenocarcinoma. KLK13 can be used as a diagnostic marker for cervical adenocarcinoma. The expression of KLK13 in HeLa cervical cancer cells is interfered with by siRNA, and the MTT cell proliferation test and the cell scratch test are respectively It showed that the proliferation and migration ability of cervical cancer cells were significantly increased compared with the control group.

Description

technical field [0001] The invention relates to the field of immune detection, in particular to the application of KLK13 in the preparation of a detection kit for diagnosing cervical adenocarcinoma. Background technique [0002] Cervical cancer is one of the most common female malignancies, ranking fourth among cancer-causing deaths among women, of which cervical squamous cell carcinoma accounts for 80%-85%, and cervical adenocarcinoma accounts for 10%-15%. With the popularization of screening for two cancers, the incidence of cervical squamous cell carcinoma has been significantly reduced, while cervical adenocarcinoma is relatively hidden due to the location of the lesion, and the existing screening methods have not changed its incidence and mortality. At present, the early detection of cervical adenocarcinoma still has the characteristics of low specificity and easy misdiagnosis and misdiagnosis. Moreover, the current research generally believes that the prognosis of cer...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/573
CPCG01N33/573G01N33/57411G01N33/57488G01N2333/96455
Inventor 顾芸储诚卓钟天鹰刘妍徐瑞朱一超
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products