Plasma metabolism micromolecular markers related to lung cancer early diagnosis and application thereof
A technology for metabolizing small molecules and early diagnosis, applied in material separation, analysis materials, measurement devices, etc., can solve the problems of high missed or misdiagnosed rate, low detection rate of markers, limited clinical application, etc., and achieve strong correlation between outcomes , Improve sensitivity and specificity, and achieve the effect of accurate judgment
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experiment example 1
[0043] One. Objectives and methods
[0044] 1. Specimen collection and processing
[0045] Collect whole blood samples of 43 healthy volunteers and 43 preoperative lung cancer patients and place them in K 2 In the EDTA blood collection tube, centrifuge at 4°C and 8000rpm for 10 minutes, take the upper layer of plasma and EP tube, and quickly place it in a cold storage at -80°C for testing. Take 20μL from the plasma samples of healthy volunteers, mix them to prepare QC samples, and store them in the same way.
[0046] 2. Main reagents
[0047] Heptadecanoic acid (content> 99.9%), methoxyamine hydrochloride (MOX, content> 99.9%), N-methyl trimethylsilyl trifluoroacetamide (MSTFA, content> 99.9%) and pyridine were purchased from Sigma-Aldrich (St. Louis, MO, USA); formic acid, methanol and acetonitrile (HPLC reagents, Merck, Germany); ultrapure water (MILIQ ultrapure water from Milipore, USA) ).
[0048] 3. Detection and identification of plasma differential metabolites
[0049] 3.1. UPL...
Embodiment 2
[0088] Example 2: Preparation of the detection kit:
[0089] Based on the plasma metabolism small molecule markers provided by the present invention, a detection kit is prepared, which contains the following components:
[0090] Standards for plasma metabolism small molecular markers: including propionic acid, aspartic acid, glucopyranoside, citric acid, allose, talose, galactopyranos, pyruvate, tryptophan, leucylpro Acid, glycerol phosphorylcholine, oleoylcarnitine, lysophosphatidylcholine (18:3), lysophosphatidylcholine (18:2), lysophosphatidylcholine (18:1), lysophosphatidylcholine Choline (22:6), lysophosphatidylethanolamine (20:0), lysophosphatidylethanolamine (18:2), tyrosylvaline;
[0091] Solvent for extracting plasma metabolites: a 1:1 ratio of methanol and acetonitrile mixed solution, 80% methanol aqueous solution (for UPLC-IT-TOF / MS sample preparation); methanol, methoxyamine hydrochloride and N-methyltrimethyl Silicon-based trifluoroacetamide (for GC-MS sample preparati...
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