Liquid chip kit for diagnosing lung cancer

A liquid phase chip and lung cancer diagnosis technology, applied in the field of medicine and biology, can solve the problems of missed diagnosis, weak specificity, and low sensitivity of patients, and achieve the effect of high accuracy.

Inactive Publication Date: 2013-11-13
HENGDIAN GRP JIAYUAN CHEM +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods are all single-indicator detection methods: the detection of a single marker for tumors always has shortcomings such as lo...

Method used

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  • Liquid chip kit for diagnosing lung cancer
  • Liquid chip kit for diagnosing lung cancer
  • Liquid chip kit for diagnosing lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Preparation of CTAG antibody, CRP and Prolactin marker detection kit for lung cancer serum samples

[0044] 1. Reagents and solutions

[0045] (1) 0.1M NaH 2 PO 4 , pH6.2: Weigh 3.5814g NaH 2 PO 4 in 90mL ddH 2 In O, adjust the pH to 6.2 with NaOH, dilute to 100mL, and filter with a 0.22um filter;

[0046] (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;

[0047] (3) 0.1M MES, 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;

[0048] (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;

[0049] (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;

[0050] (6) 1%PBSB: 0.1%PBS containing 0.02%Proclin300, 0.22um membrane filtration;

[...

Embodiment 2

[0100] Embodiment 2 Liquid phase chip kit of the present invention is used for the detection of lung cancer

[0101](1) Take out all the reagents before use, and let them equilibrate to room temperature;

[0102] (2) Take 2-plex Halo Tag (O 2 ) Amine ligand-coupled microspheres, vortexed for 20s, ultrasonicated for 20s, according to 2500 microspheres per well, take an appropriate amount in 500ul of 1%PBSB;

[0103] (3) Protect from light and seal for 1 hour;

[0104] (4) 14000g, centrifuge for 4min, absorb the supernatant and add the corresponding protein, among them, CTAG (Creative Biomart, Cat. No. CTAG1B-1512H), Halo each 0.2ug / well, supplemented with 1% PBSB to a final volume of 500ul;

[0105] (5) Incubate at room temperature in the dark for 1 hour;

[0106] (6) After vortexing and ultrasonicating the above microspheres for about 20s, dispense them into 96-well plates, 10ul / well;

[0107] (7) Prepare CRP (R&D Systems, product number 1229006), Prolactin (R&D Systems, p...

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Abstract

The invention discloses a liquid chip kit for diagnosing lung cancer. The kit comprises microballs coated with capturing antibodies, detecting antibodies labeled by biotin, microballs coupled with Halotag Amine (O2) ligand, antigen proteins, streptavidin-phycoerythrin and anti-human immune globulin G labeled by biotin; wherein the capturing antibodies contain at least one component, which is selected from the following capturing antibodies: CRP, Prolactin, CEA, NSE, CK19, Axl and ADAM8; the detecting antibodies contain at least one component, which is selected from the following detecting antibodies: CRP, Prolactin, CEA, NSE, CK19, Axl and ADAM8, and are corresponding to the capturing antibodies; the antigen proteins contain at least one component, which is selected from following components: p62, CTAG, p53 and CAGE. Microballs with different antibodies or antigen proteins have different color codes. The liquid chip kit carries out detections of antigen and autoantibody at the same time, and the precision rate is high.

Description

technical field [0001] The invention relates to the field of medicine and biology, in particular to a liquid phase chip kit for lung cancer diagnosis. Background technique [0002] Lung cancer is currently the malignant tumor with the highest morbidity and mortality worldwide. In the past 20 years, due to the vigorous promotion of smoking cessation, the incidence of male lung cancer in Western countries such as Europe and the United States has begun to decline. But the incidence of lung cancer in women continues to rise. my country is a big country in the production and sales of cigarettes, regardless of men or women. The incidence of lung cancer has continued to rise, especially in women. Clinical studies have shown that the cure rate of carcinoma in situ is close to 100%. The 5-year survival rate of patients with stage I lung cancer is 60% to 90%, while the 5-year survival rate of patients with stage IIIb and IV lung cancer is only 5% to 20%. It suggested that early d...

Claims

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Application Information

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IPC IPC(8): G01N33/574
Inventor 王孝举吴国君马胜林汪成发夏冰陈雪琴
Owner HENGDIAN GRP JIAYUAN CHEM
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