Application of serum biomarkers in the preparation of diagnostic reagents for issu
A biomarker and serum technology, applied in the field of serum biomarker combinations, can solve the problems of increasing gastrointestinal and respiratory tract infections in patients, and achieve the effect of improving treatment effect, improving patient prognosis, and stabilizing results
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Embodiment 1
[0030] Example 1: Screening characterization of serum differential metabolites between IS patients and ISSU patients
[0031] 1. Objects and Methods
[0032] 1. Sample collection and processing
[0033] The study was approved by the hospital ethics committee, and the patients or their families signed informed consent. Serum samples from 33 IS patients and 15 ISSU patients were collected and stored at -80°C for cryopreservation. Take 20 μL from each sample to be tested, mix well to prepare QC samples, and store them under the same conditions.
[0034] 2. Standards and Reagents
[0035] Stable isotope-labeled standards D1-leucine (content 99%) and heptadecanoic acid (content 99.9%) were purchased from Sigma-Aldrich Company in the United States, and D5-cholic acid (content 98%) was purchased from Cambridge Isotope Laboratories Company in the United States. Dansyl chloride (Dns-Cl, content 98%) was purchased from Shanghai Bailingwei Company; dansylpiperazine (Dns-PP) was prepa...
Embodiment 2
[0065] Example 2: Construction of a ROC curve for verifying the ability of 6 differential metabolites to diagnose IS and ISSU
[0066] Receiver operating curve (ROC) method was used for verification, and the ability to diagnose and distinguish IS and ISSU patients was judged by the expression levels of differential metabolites in the serum of IS and ISSU patients. The results showed that the six differential metabolites, glycine, phenylalanine, ornithine, palmitoleic acid, arachidonic acid, and oleic acid, had a strong ability to diagnose and distinguish between IS and ISSU patients, and the area under the ROC curve ( AUC) are greater than 0.7, which has clinical diagnostic significance; when used in combination for diagnosis, with the increase of the number of combinations, the AUC further increases, the highest when all 6 are combined, the AUC reaches 0.986, and the sensitivity and specificity are 93.3% and 93.9% respectively %. See Tables 2-4 for single and any combination...
Embodiment 3
[0075] Embodiment 3: the preparation of detection kit:
[0076] Based on the serum biomarkers provided by the present invention, a detection kit is prepared, which contains the following components:
[0077] Serum biomarker standards: glycine, phenylalanine, ornithine, palmitoleic acid, arachidonic acid, oleic acid;
[0078] Detection reagents: dansyl chloride, dansyl piperazine, 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate, ethyl acetate, acetone , acetonitrile, methanol, formic acid (for sample preparation, derivatization reaction and LC-MS / MS determination);
[0079] Internal standard: Stable isotope labeled standards D1-leucine, D5-cholic acid and heptadecanoic acid.
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