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Detecting method for lung cancer characteristic metabolite fingerprint spectrum in urine

A technology of metabolites and fingerprints, which is applied in the field of detection of fingerprints of metabolites characteristic of lung cancer in urine, achieves high sensitivity and specificity, meets the requirements of simplicity and effectiveness

Active Publication Date: 2013-04-10
CHINA TOBACCO ZHEJIANG IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main treatment of lung cancer depends on early diagnosis and treatment, and currently there is no convenient and effective method for screening lung cancer

Method used

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  • Detecting method for lung cancer characteristic metabolite fingerprint spectrum in urine
  • Detecting method for lung cancer characteristic metabolite fingerprint spectrum in urine
  • Detecting method for lung cancer characteristic metabolite fingerprint spectrum in urine

Examples

Experimental program
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Embodiment 1

[0029] The detection method of embodiment 1 urine

[0030] Pipette 100 μl of urine sample into a 1.5ml centrifuge tube, add 10 μl of urease (used to remove urea in urine, urea can interfere with the dehydration and derivatization of other water-soluble metabolites in urine) aqueous solution (80mg / ml), After mixing evenly, incubate in a water bath at 37°C for 2 hours to remove urea contained in the urine, add internal standard 10 μL pentapentyl alcohol aqueous solution (0.3 mg / ml) and 200 μL acetone, vortex for 1 min, and after fully mixing, sonicate in an ice bath for 15 min , centrifuge (10000r / min) for 10min, draw 200μL of the supernatant into a GC sample bottle, centrifuge and concentrate at 35°C for 4h and evaporate the solvent. Add 50 μL of methoxyaminopyridine solution (15 mg / ml), mix well, oximate at 70°C for 1 h, and then add derivatization reagent (MSTFA:TMCS=100:1, V / V, MSTFA is N-methyl-N- (trimethylsilane)trifluoroacetamide, TMCS is trimethylchlorosilane) 50μL, mi...

Embodiment 2

[0031] Example 2 The method of bioinformatics finds the differential metabolites of lung cancer and healthy controls in smoking and non-smoking populations

[0032]A total of 128 urine samples were collected, including 49 lung cancer patients (31 of which were smokers) and 79 healthy controls (36 of which were smokers). Each urine sample was detected using the urine detection method in Example 1, and finally the obtained data was analyzed by bioinformatics using ZJU-PDAS software (developed by Cancer Institute of Zhejiang University). 9 / 10 samples were randomly selected from the smoking population (including lung cancer patients and healthy controls) as the training set, and the remaining 1 / 10 samples were used as the test set. Use the training set to find the differential metabolites between lung cancer patients and healthy controls, and use the support vector machine method to select 7 metabolites from the lung cancer metabolite fingerprints of smokers, and use this map to e...

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Abstract

The invention discloses an establishing method for a lung cancer characteristic metabolite fingerprint spectrum in urine. The gas chromatography and mass spectrum combined technology is adopted to detect metabolic profiles of urine samples of lung cancer patients and healthy donors, and is combined with the biological method of a support vector machine to build a smoker lung cancer metabolite fingerprint spectrum and a non-smoker lung cancer metabolite fingerprint spectrum, wherein the smoker lung cancer metabolite fingerprint spectrum comprises oxalic acid, phosphoric acid, uracil, threonine, 5-pidolic acid, citric acid and galactose or comprises phosphoric acid, uric acid, citric acid and oxalic acid; and the non-smoker lung cancer metabolite fingerprint spectrum comprises alanine, oxalic acid, phosphoric acid, uracil, serine, threonine, 5-pidolic acid, ribose, aconitate, citric acid, galactose, tyrosine, palmitic acid and stearic acid. The method can detect only needing the urine samples, satisfies the requirements of effectiveness, simplicity and feasibility in the operation aspect, and is suitable for large-scale application.

Description

technical field [0001] The invention relates to the field of urine detection, in particular to a method for detecting fingerprints of lung cancer characteristic metabolites in urine. Background technique [0002] Lung cancer is the most common primary malignant tumor of the lung. The vast majority of lung cancer originates from the bronchial mucosal epithelium, so it is also called bronchial lung cancer. It is the tumor with the highest morbidity and mortality in the world. Its overall 5-year survival rate is about 15% in the United States and about 8% in China and European countries. However, the 5-year survival rate of advanced distant metastatic cases is only 2%. At present, the main treatment method is early detection and early treatment. The occurrence and development of lung cancer are usually hidden, and 7%-10% of the cases have no clinical manifestations, and they are already at an advanced stage when they are discovered. There is still a lack of a simple and effe...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 储国海周国俊黄芳芳余捷凯胡集祎郑树范骁辉陆晓燕
Owner CHINA TOBACCO ZHEJIANG IND
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