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Detection method for activity of kallikrein 7 in blood and application of kallikrein 7 as a serum marker for diagnosing pancreatic cancer

A technology for detecting kallikrein and activity, applied in the field of medicine

Inactive Publication Date: 2018-12-04
YICHANG CENT PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there is no effective serum marker for clinical diagnosis and treatment of pancreatic cancer

Method used

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  • Detection method for activity of kallikrein 7 in blood and application of kallikrein 7 as a serum marker for diagnosing pancreatic cancer
  • Detection method for activity of kallikrein 7 in blood and application of kallikrein 7 as a serum marker for diagnosing pancreatic cancer

Examples

Experimental program
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example 1

[0018] Example 1: The expression level of KLK7 in pancreatectomy tissues of patients with pancreatic ductal carcinoma was detected by immunohistochemical method. From October 2015 to October 2016, pancreatic cancer tissues and paracancerous tissues were collected from 46 patients who were diagnosed with pancreatic cancer by pathology at the Department of Hepatobiliary and Pancreatic Surgery of Yichang Central People's Hospital. Immediately after resection, the specimens were fixed in 10% neutral buffered formalin, dehydrated, transparent, embedded in paraffin, and made into 4 μm thick sections for immunohistochemical staining. Immunohistochemical staining was detected using the EnVison two-step method according to the kit instructions.

[0019] Detection method: EnVison two-step method

[0020] 1) Place the paraffin sections in an oven at 67°C, and bake the sections for 2 hours;

[0021] 2) Paraffin sections were dewaxed, hydrated, and rinsed with tap water;

[0022] 3) Put...

example 2

[0030] Example 2: Detection of the activity of KLK7 protein in the serum of patients with pancreatic cancer, patients with pancreatic cystadenoma, patients with pancreatitis and normal people. Sera were collected from patients who visited the Department of Hepatobiliary and Pancreatic Surgery, First Clinical Medical College of China Three Gorges University from February 2016 to October 2016, 32 patients with pancreatic cancer (before surgery), 32 patients with pancreatic cystadenoma, and patients with pancreatitis Serum from 32 cases, and another 32 normal volunteers with serum collected randomly. The activity of KLK7 in serum was detected by FRET technology (Enzolifescience, Kallikrein-7 (human), ELISA Kit, ADI-900-218-0001; McaRPKPVENvalWRK(Dnp)NH2, R&D systems, 2624-SE).

[0031] Preparation of items and equipment before testing

[0032] 1) All blood samples to be tested should be placed in a refrigerator at 4°C for 12 hours as required, then centrifuged and the supernatan...

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Abstract

The invention provides a detection method for activity of kallikrein 7 in blood. The method includes the following steps: subjecting collected to-be-detected blood samples to stand in a refrigerator at 4 DEG C for 12 hours, and then taking a supernatant by centrifugation and storing the supernatant in a refrigerator at -80 DEG C; before detection, according to a ratio of 1:5, diluting serum with 1*PBS, and subjecting the serum to split charging and a to-be-detected experiment; distributing an enzyme-linked plate with 96 holes, adding a to-be-detected serum sample of 50 [mu]L, gently shaking, and incubating for 2 h; removing a liquid in the holes, adding a wash buffer, washing the holes 3 times, placing in a shaker for 5 minutes after the buffer is added each time, flatly paving absorbent paper on a tabletop, and tapping the enzyme plate to cleanly remove the liquid in the holes; adding KLK7 FRET fluorogenic substrate McaRPKPVENvalWRK (Dnp) NH2 to each of the holes, and incubating for 30 minutes at 37 DEG C on the shaker; and performing detection at an excitation light wavelength of 320 nm and an emission light wavelength of 405 nm in a fluorescence enzyme-labeled instrument.

Description

technical field [0001] The invention relates to the field of medicine, in particular to the detection of activity of Kallikrein 7 in blood and its application in clinical diagnosis of pancreatic cancer. Background technique [0002] Pancreatic cancer is a highly malignant tumor of the digestive system, and its fatality rate accounts for about 83% of the annual incidence. In the 1980s, the 5-year survival rate of pancreatic cancer was about 3%, and it reached 7% in the early 21st century. Due to the insidious onset and the unique anatomical structure and biological characteristics of the pancreas, 60% of pancreatic cancer patients have distant metastases at the time of diagnosis, and are not sensitive to traditional chemotherapy and radiotherapy, so the prognosis is extremely poor, with a median survival of The period is only 6 to 9 months. Pancreatic ductal adenocarcinoma is the most common pathological type of pancreatic cancer, and pathological types such as cystadenocar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
CPCG01N33/57438G01N33/57492G01N33/6893
Inventor 谭潇刘森郑军李林张丁
Owner YICHANG CENT PEOPLES HOSPITAL