Gluconolactone and pyroglutamic acid as macrosomia auxiliary diagnostic marker and application thereof
A technology of gluconolactone and pyroglutamic acid, which is applied in the field of metabolic small molecule marker combination, can solve the problems of complex experimental steps and low sensitivity, and achieves improved sensitivity and specificity, high sensitivity and specificity, and ease of use. The effect of detection
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Embodiment 1
[0093] Embodiment 1 Research Object Selection and Grouping Basis
[0094] The present inventor collected blood samples from mothers of macrosomia patients and normal children who met the requirements from the Nanjing Maternal and Child Health Hospital Affiliated to Nanjing Medical University (see figure 1 , figure 2 ). Weigh the newborn after birth, and if the weight is equal to or greater than 4kg within 1 hour after birth, it is diagnosed as "macrosomia". In the first stage, 97 samples of pregnant women who met the requirements were randomly included, 15 of which were macrosomia; in the second stage, 30 samples of pregnant women were included, 15 of which were compared with 15 cases of macrosomia, as the blood metabolism of small molecular organisms during pregnancy of macrosomia. Screening subjects for markers. Specific sample classification criteria are as follows:
[0095] first stage screening
[0096] A total of 97 pregnant women were randomly included.
[0097] ...
Embodiment 2
[0110] Example 2 UPLC-MS metabolomics biomarker screening for macrosomia
[0111] 1. Sample pretreatment
[0112] 1. Centrifuge fresh blood during pregnancy at 3000rpm for 5min in a centrifuge, take 100μl of the supernatant and dispense it into clean 1.5ml EP tubes.
[0113] 2. Add 50 μl of serum to 50 μl of water and 300 μl of methanol (reagent A) to precipitate protein.
[0114] 3. Aspirate the supernatant, blow dry with nitrogen and then vacuum dry.
[0115] 4. Reconstitute the dry matter (Reagent D).
[0116] 2. Instrument testing
[0117] 1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.
[0118] 2. Liquid phase conditions:
[0119] The liquid chromatography column is a Hypersil GOLD C18 column (100mm×2.1mm, particle size 1.9μm, ThermoScientific, Germany), and the column temperature is 40°C.
[0120] The mobile phases employed were (A) 0.1% formic acid in water (reagent ...
Embodiment 3
[0132] Example 3 Stability Analysis of Small Molecules in Serum Metabolism
[0133] The method of Example 2 was used to evaluate the stability of the combined serum gluconolactone and pyroglutamic acid levels of the mother during pregnancy (the interval was 2 weeks). The results showed that the measured levels of gluconolactone and pyroglutamic acid in serum were stable ( Figure 4 ), with properties as diagnostic / monitoring markers.
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