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Mass spectrum model for detecting liver cancer serum characteristic protein and method for preparation

A technology of characteristic protein and mass spectrometry model, which is applied in the field of protein fingerprint detection to achieve high sensitivity and specificity, reasonable and feasible construction method, and the effect of reducing the fatality rate

Inactive Publication Date: 2005-06-01
THE 306TH HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the detection of serum characteristic proteins of liver cancer using SELDI-TOF-MS technology in China so far.

Method used

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  • Mass spectrum model for detecting liver cancer serum characteristic protein and method for preparation
  • Mass spectrum model for detecting liver cancer serum characteristic protein and method for preparation

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Draw 2 mL of venous blood from the patient to be tested without anticoagulation. Place at room temperature for 30 min to 1 h, centrifuge at room temperature at 2500 rpm for 5 min, take aliquots of serum and store in a -70°C refrigerator. During the experiment, the samples were taken out from the -70°C refrigerator, thawed in ice, and centrifuged at 10,000 rpm for 2min at 4°C. Take 3 μL of centrifuged serum sample, add 6 μL of U9 treatment solution (9M urea, 2% CHAPS, 1% DTT, 50mM Tris-CL, pH9.0), mix well, shake in ice bath for 30min, take it out, add 108μl of binding buffer (100mmol / LNaAc, PH4.0), mix immediately. Put the WCX2 chip into the bioprocessor, add 200 μl of binding buffer to each well, shake and wash at room temperature for 2 times, each time for 5 minutes, and spin dry. Add 100 μl sample mixture to each well, shake and incubate for 1 h, shake off the sample, wash twice with 200 μl elution buffer (100 mmol / L NaAc, pH 4.0) at room temperature for 5 min each...

Embodiment 2

[0046] Draw 2 mL of venous blood from the patient to be tested without anticoagulation. Place at room temperature for 30 min to 1 h, centrifuge at room temperature at 2500 rpm for 5 min, take aliquots of serum and store in a -70°C refrigerator. During the experiment, the samples were taken out from the -70°C refrigerator, thawed in ice, and centrifuged at 10,000 rpm for 2min at 4°C. Take 3 μL of centrifuged serum sample, add 6 μL of U9 treatment solution (9M urea, 2% CHAPS, 1% DTT, 50mM Tris-CL, pH9.0), mix well, shake in ice bath for 30min, take it out, add 108μl of binding buffer (100mmol / LNaAc, PH4.0), mix immediately. Put the WCX2 chip into the bioprocessor, add 200 μl of binding buffer to each well, shake and wash at room temperature for 2 times, each time for 5 minutes, and spin dry. Add 100 μl sample mixture to each well, shake and incubate for 1 h, shake off the sample, wash twice with 200 μl elution buffer (100 mmol / L NaAc, pH 4.0) at room temperature for 5 min each...

Embodiment 3

[0048] Draw 2 mL of venous blood from the patient to be tested without anticoagulation. Incubate at 37°C for 20-30min, centrifuge at room temperature at 2500rpm for 5min, take aliquots of serum and store in -70°C refrigerator. During the experiment, the samples were taken out from the -70°C refrigerator, thawed in ice, and centrifuged at 10,000 rpm for 2min at 4°C. Take 3 μL of centrifuged serum sample, add 6 μL of U9 treatment solution (9M urea, 2% CHAPS, 1% DTT, 50mM Tris-CL, pH9.0), mix well, shake in ice bath for 30min, take it out, add 108μl of binding buffer (100mmol / LNaAc, PH4.0), mix immediately. Put the WCX2 chip into the bioprocessor, add 200 μl of binding buffer to each well, shake and wash at room temperature for 2 times, each time for 5 minutes, and spin dry. Add 100 μl sample mixture to each well, shake and incubate for 1 h, shake off the sample, wash twice with 200 μl elution buffer (100 mmol / L NaAc, pH 4.0) at room temperature for 5 min each time, and dry; th...

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Abstract

The present invention is mass spectral model for detecting liver cancer serum characteristic protein and its preparation process, and belongs to the field of protein fingerprint detecting technology. The present invention screens 17 characteristic proteins including 6 up regulation proteins and 11 down regulation proteins, selects two or more proteins out of the 17 characteristic proteins and establishes two or more stage classification tree with the clinical peak values of the proteins to constitute the detection model for detecting the liver cancer serum characteristic protein. The present invention lays foundation for further discovering new liver cancer biological mark, and the model has over 90 % specificity, sensitivity and positive predicting rate in liver cancer diagnosis.

Description

technical field [0001] The invention belongs to the technical field of detection of protein fingerprints, in particular to a detection method for serum protein fingerprints of liver cancer. Background technique [0002] Liver cancer is one of the most common malignant tumors, with a high mortality rate and a serious threat to people's health. About 110,000 people die of liver cancer every year in China, accounting for 45% of the world's liver cancer deaths. Early diagnosis and early treatment are the key to improve the survival rate of patients. However, the sensitivity and specificity of current diagnostic methods are low, which has become a bottleneck in the diagnosis and treatment of liver cancer. The high morbidity and high mortality rate of liver cancer make people constantly seek an effective early diagnosis method. It is far from enough to use AFP protein as a serological diagnostic index clinically in terms of sensitivity and specificity. Once the space-occupying ...

Claims

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Application Information

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IPC IPC(8): G01N30/00G01N33/68
Inventor 张建中郑燕华冯凯邹德威
Owner THE 306TH HOSPITAL OF PLA
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