Urine biomarker for early diagnosis of angiostrongyliasis cantonensis as well as screening method and application of urine biomarker
A biomarker, Angiostrongylus nematode technology, applied in the field of biomedicine, can solve the problems that have not been seen before, and achieve the effect of accurate reflection and high specificity
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Embodiment 1
[0037] Example 1 Establishment of Angiostrongylus cantonensis Infected Mouse Model and Sample Collection and Processing
[0038] 1. Establishment of mouse model infected with Angiostrongylus cantonensis
[0039] Healthy SPF level BALB / c mice aged 6-8 weeks were randomly divided into 10 groups: Angiostrongylus cantonensis infection group for 1 day, normal control group for 1 day, Angiostrongylus cantonensis infection group for 3 days, normal control group for 3 days, Guangzhou Angiostrongylus infection group for 7 days, normal control group for 7 days, Angiostrongylus cantonensis infection group for 14 days, normal control group for 14 days, Angiostrongylus cantonensis infection group for 21 days and normal control group for 21 days, 10 mice in each group . The normal control group was not treated, and the mice in the infection group were infected according to the conventional method: crush the smooth biumbilical snail containing the stage III larva of Angiostrongylus cantonen...
Embodiment 2
[0042] Example 2 Analysis of urine metabolites in mice infected with Angiostrongylus cantonensis by ultra-high performance liquid chromatography-tandem mass spectrometry
[0043] 100 samples obtained in Example 1 were analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry, and the specific analysis conditions were as follows:
[0044] 1. Instruments and equipment
[0045] Liquid Chromatography: Waters ACQUITY UPLC-I Class System
[0046] Mass spectrometry: Waters SYNAPT G2-Si HDMS
[0047] Chromatographic column: ACQUITY UPLC C18 BEH chromatographic column (2.1×100mm, 1.7μm)
[0048] 2. Liquid chromatography conditions
[0049] Mobile phase: A-H 2 O (0.1% formic acid), B-acetonitrile (0.1% formic acid)
[0050] Flow rate: 400μL / min; Column temperature: 40℃; Injection volume: 0.600μL
[0051] Gradient elution program: 0-1min(1%B), 1-4min(1%-10%B), 4-5min(10%B), 5-9min(10%-50%B), 9-11min (50%-95%B), 11-12.5min (95%B), 12.6-15min (5%B).
[0052]...
Embodiment 3
[0054] Example 3 Analysis of urine metabolites in mice infected with Angiostrongylus cantonensis by ultra-high performance liquid chromatography-tandem mass spectrometry
[0055] 100 samples obtained in Example 1 were analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry, and the specific analysis conditions were as follows:
[0056] 2. Instruments and equipment
[0057] Liquid Chromatography: Waters ACQUITY UPLC-I Class System
[0058] Mass spectrometry: Waters SYNAPT G2-Si HDMS
[0059] Chromatographic column: ACQUITY UPLC C18 BEH chromatographic column (2.1×100mm, 1.7μm)
[0060] 3. Liquid Chromatography Conditions
[0061] Mobile phase: A-H 2 O (0.1% formic acid), B-acetonitrile (0.1% formic acid)
[0062] Flow rate: 400μL / min; Column temperature: 45℃; Injection volume: 0.600μL
[0063] Gradient elution program: 0-1min(1%B), 1-4min(1%-10%B), 4-5min(10%B), 5-9min(10%-50%B), 9-11min (50%-95%B), 11-12.5min (95%B), 12.6-15min (5%B).
[0064]...
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