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Method for detecting N-glycome log (P9/P4) in serum and detecting system and application thereof

A detection system, serum technology, applied in the field of clinical medical diagnostics, can solve problems such as adoption

Active Publication Date: 2009-11-11
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for a long time, glycosylation analysis has been difficult to be adopted in clinical research due to technical bottlenecks

Method used

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  • Method for detecting N-glycome log (P9/P4) in serum and detecting system and application thereof
  • Method for detecting N-glycome log (P9/P4) in serum and detecting system and application thereof
  • Method for detecting N-glycome log (P9/P4) in serum and detecting system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Analysis of N-glycan groups in the normal control group, liver fibrosis group and HCC patient group Collected primary liver cells from January 2007 to April 2008 in Dongfang Hepatobiliary Surgery Hospital Affiliated to Second Military Medical University The serum of 145 patients with cancer. Serum samples from 128 patients with liver fibrosis in Renji Hospital of Shanghai Jiaotong University were collected during the same period. All the above patients met the following inclusion criteria: ①all HCC patients with HBV infection; ②Patients’ postoperative pathological specimens were all diagnosed as HCC patients or patients with liver fibrosis by pathologists; ③Human immunodeficiency virus (human immunodeficiency virus, HIV) was excluded , hepatitis C virus (HCV), hepatitis D virus (HDV), hepatitis E virus HEV, and syphilis other than hepatitis B virus infection; ④ Autoimmune liver disease, alcoholic liver disease, drug-induced liver disease, and Wilson disease ...

Embodiment 2

[0059] Embodiment 2: Detect patient's serum N sugar log (P9 / P4)

[0060] Specific experimental steps:

[0061] 1) Blood was drawn from patients 1, 2, 3 and 4, and serum was collected.

[0062] 2) Oligosaccharide release: 2 μl serum + 2 μl buffer solution (10 mM NH4HCO3, 5% SDS) + 3 μl water; heat at 95°C for 5 minutes, place at 4°C for 15 minutes; add 3 μl PNGaseF (2.2U / μl); incubate at 37°C for 3 Hours, cool at 4°C; add 100 μl of water, mark tube D.

[0063] 3) Labeling of oligosaccharides: take 20 μl (solution in tube D), dry at 60°C for 3 hours; add 2 μl of labeling solution (20mM APTS:1M NaCNBH3=1:1); incubate at 37°C for 16 hours; add 200μl of water to terminate the reaction, Marked as L tube.

[0064] 4) Oligosaccharide desialic acid: 2 μl (L tube solution) + 3 μl desialidase (2 mU); incubate overnight at 37°C; shake and mix with 35 μl water, and mark it as a DE tube.

[0065] 5) Detection and analysis: 10 μl (DE tube solution) was used for fragment analysis with an ...

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Abstract

The invention belongs to the diagnostics field of clinical medicine and aims at providing a method for detecting N-glycome log (P9 / P4) in serum so as to overcome defects of prior art that both sensitivity and specificity of AFP detection and supersonic inspection which are adopted to screen high risk group of HCC which is the fifth common malignancy in the world are so low that HCC cannot be detected and treated early. The method is based on DSA-FACE of a DNA sequencer to detect distribution of glycan connected by N-glycosidic bond on glycosidoprotein in serum so as to calculate value of log (P9 / P4). The invention also provides a corresponding detecting system and applies the detecting method and the detecting system to the early screening and diagnosis of HCC, especially to the early screening and diagnosis of tendency of HCC and hepatic fibrosis relevant to HBV inflection. Compared with the AFP detection, the method provided by the invention can raise the specificity and accuracy respectively by 16 percent and 8 percent and has the similar sensitivity.

Description

technical field [0001] The invention belongs to the field of clinical medical diagnostics, and in particular relates to a method for detecting serum N-glycan log (P9 / P4), a detection system and an application thereof. Background technique [0002] Primary hepatocellular carcinoma (HCC) is the fifth most common malignant tumor in the world, and it is also one of the tumors with the fastest increasing incidence and the highest mortality rate in recent years. In my country, the high incidence of chronic liver disease and liver cirrhosis is caused by the high infectivity of HBV. According to research reports, these high-risk groups have a 10-25% chance of developing HCC in their lifetime. The onset of HCC is mostly insidious, and there are usually no clinical symptoms in the early stage, or only the symptoms and signs of hepatitis, so it is difficult to detect early. Once the typical clinical manifestations appear, such as pain in the liver area, weight loss, jaundice or ascit...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N27/447G01N33/66G01N33/574
Inventor 高春芳房萌赵云鹏王皓顾星周飞国赵金艳季君
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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