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Liquid chip kit for detecting lung cancer autoantibody

A liquid phase chip and autoantibody technology, applied in the field of medicine and biology, can solve the problems of low sensitivity and accuracy, and achieve the effect of rapid detection and high detection accuracy

Inactive Publication Date: 2013-11-06
HANGZHOU FIRST PEOPLES HOSPITAL +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This fully shows that the single detection of tumor markers has high specificity, but low sensitivity and accuracy; after combined detection, although the specificity is reduced, the sensitivity and accuracy are greatly improved

Method used

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  • Liquid chip kit for detecting lung cancer autoantibody
  • Liquid chip kit for detecting lung cancer autoantibody
  • Liquid chip kit for detecting lung cancer autoantibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The preparation of embodiment 1 liquid phase chip kit

[0033] Reagents and Solutions

[0034] (1) 0.1M NaH 2 PO 4 , pH6.2: Weigh 3.5814g NaH 2 PO 4 in 90mL ddH 2In O, adjust the pH to 6.2 with NaOH, dilute to 100mL, and filter with a 0.22um filter;

[0035] (2) 0.05M MES, pH5.0: weigh 0.976g MES in 90mL ddH 2 In O, adjust the pH to 5.0 with NaOH, dilute to 100mL, and filter with a 0.22um filter;

[0036] (3) 0.1M MES (2-(N-morpholino)ethanesulfonic acid), pH6.0: weigh 1.952g MES in 90mL ddH 2 In O, adjust the pH to 6.0 with NaOH, dilute to 100mL, and filter with a 0.22um filter;

[0037] (4) 0.1M MES, pH4.5: weigh 1.952g MES in 90mL ddH 2 In O, adjust the pH to 4.5 with NaOH, dilute to 100mL, and filter with a 0.22um filter;

[0038] (5) PBS: NaCl, 137mM; KCl, 2.7mM; NaCl 2 HPO 4 , 8.1mM; KH 2 PO 4 , 1.5mM; pH7.2-7.4, 0.22um membrane filtration;

[0039] (6) 1%PBSB: 0.1%PBS containing 0.02%Proclin300, 0.22um membrane filtration;

[0040] (7) 0.1%PBST: 0...

Embodiment 2

[0125] Example 2 Lung cancer detection test

[0126] 1. Detection method

[0127] (1) Take out all the reagents before use, and let them equilibrate to room temperature;

[0128] (2) Take out the serum of patients (suffering from lung cancer) and healthy people at -80°C, place them on ice to melt, vortex and mix well, take V-type 96-well hemagglutination plate (96-well plate) and dilute the serum 200 times, Mix the tip of the pipette up and down, try not to generate air bubbles;

[0129] (3) Take the 4 kinds of microspheres coupled with antigenic protein in Example 1, vortex and mix for 20s, sonicate for 20s, take an appropriate amount according to 2500 microspheres per well, and put it in 1000ul1%PBSB, vortex and mix for 20s, sonicate for 20s, The final volume is 10ul / well;

[0130] (5) Turn on the plate washer---Prime----rinse (channel2)----Prime---place the 96-well plate---Run5: MAGX3;

[0131] (6) Immediately add serum diluted 200 times, 30ul / well;

[0132] (7) Wrap t...

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Abstract

The invention discloses a liquid chip kit for detecting a lung cancer autoantibody. The kit includes: microspheres coupling antigen proteins, a biotin labeled second antibody, streptavidin-phycoerythrin, a reaction buffer and a dilution buffer solution. The antigen proteins are at least two of p62, NY-ESO-1, p53 and CAGE, wherein the microspheres coupling different antigen proteins have different color codes. The second antibody is anti-human immunoglobulin G. The liquid chip kit for detecting the lung cancer autoantibody provided in the invention realizes joint detection of four antibodies in serum on a liquid chip technology platform, and takes the p62 antibody, the NY-ESO-1 antibody, the p53 antibody and the CAGE antibody as marks, has high detection accuracy, and provides a new idea for early diagnosis of lung cancer.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a liquid-phase chip kit for detecting autoantibodies of lung cancer. Background technique [0002] In recent years, the incidence of cancer in my country has shown an obvious upward trend, accounting for more than 20% of the causes of death, ranking first among all causes of death. The World Health Organization predicts that by 2020, there will be 20 million new cancer cases and 12 million deaths, most of which will occur in developing countries. If no effective prevention and control measures are taken, it is estimated that by 2020, the total number of new cancers and cancer deaths in my country will reach about 3 million, and the total number of patients will reach 6.6 million. [0003] Lung cancer is currently the malignant tumor with the highest morbidity and mortality worldwide. In the past 20 years, due to vigorous promotion of smoking cessation, the incidence of male lung ...

Claims

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Application Information

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IPC IPC(8): G01N33/574
Inventor 马胜林王孝举吴国君夏冰陈雪琴王闻哲
Owner HANGZHOU FIRST PEOPLES HOSPITAL
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