Protein markers of gestational diabetes mellitus in urine and use in early diagnosis

A gestational diabetes and protein technology, which is applied in disease diagnosis, mass spectrometry, biological testing, etc., can solve the problems of sensitivity and specificity to be considered, unrecognized, and poor specificity, and achieve the goal of overcoming antibody specificity and titer Limited, high selectivity, good reproducibility

Active Publication Date: 2019-09-27
安肽和(杭州)医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the poor specificity of these indicators, and they are only screened from known mature indicato...

Method used

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  • Protein markers of gestational diabetes mellitus in urine and use in early diagnosis
  • Protein markers of gestational diabetes mellitus in urine and use in early diagnosis
  • Protein markers of gestational diabetes mellitus in urine and use in early diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1: Detection of gestational diabetes-related proteins in urine

[0092] We screened urine for gestational diabetes-associated proteins using Data independent acquisition (DIA).

[0093] Materials and Reagents

[0094] 1) Instrument: Orbitrap Fusion Lumos Tribrid mass spectrometer (Thermo Scientific Company).

[0095]2) Main reagents: trypsin (Promega Company); C18 solid-phase extraction cartridge (3CC, 60 mg, Waters Company); C18 reverse-phase chromatographic column (4.6 mm×250 mm, C18, 3 μm, Waters Company).

[0096] 3) Samples: urine from 15 patients with gestational diabetes (6-10 weeks of pregnancy) and 15 normal controls from Peking Union Medical College Hospital.

[0097] 1.1 Collection of human urine samples and enrichment of urine protein

[0098] Collect fasting morning urine and centrifuge at 5000g for 30min to remove sediment. The supernatant was precipitated with ethanol overnight at 4 to precipitate the protein. Reconstitute the protein pellet ...

Embodiment 2

[0105] Example 2: Mass spectrometric detection of urine protein NOV protein homologues, peroxide reductase-5, and hepatocyte growth factor-like proteins.

[0106] In order to better detect the application of these differential proteomic analysis results in further larger clinical samples, we used targeted mass spectrometry to monitor the differential proteins identified. Targeted analysis (parallel reaction) monitoring is a target mass spectrometry quantitative analysis technology based on the secondary mass spectrometry signal. Compared with the traditional selective reaction monitoring technology, it does not need to pre-design the parent ion / product ion pairing information of the targeted protein, saving Experimental design and operating time; and higher selectivity, better sensitivity, better reproducibility, and stronger anti-interference ability in complex backgrounds. Compared with immunization methods, it is no longer subject to commercial antibodies, and overcomes the...

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Abstract

The present invention relates to protein markers of gestational diabetes mellitus in urine and the use in early diagnosis. In particular, the present invention relates to the use of an identification reagent selected from the following proteins for preparing reagents for early diagnosis of gestational diabetes mellitus: protein NOV homolog, Peroxiredoxin-5 (PRDX-5), Hepatocyte growth factor-like protein (HFGL) and the combination thereof. The protein NOV homolog, the Peroxiredoxin-5 and the Hepatocyte growth factor-like protein and the combination thereof have good clinical application prospects for early diagnosis of gestational diabetes mellitus.

Description

[0001] field of invention [0002] The present invention relates to the field of biotechnology, more specifically to a subject sample, such as a protein marker of gestational diabetes in urine and its use in early diagnosis. [0003] Background of the invention [0004] Gestational diabetes mellitus (GDM) refers to abnormal glucose tolerance that occurs or is first discovered during pregnancy. According to different populations and diagnostic criteria, the highest incidence of GDM has reached 18%. Compared with other races, Asian women are more likely to develop GDM during pregnancy. GDM has adverse effects on the health of fetuses and pregnant women, and may cause fetal arrest, premature delivery, dystocia, infection, macrosomia, neonatal jaundice, neonatal hypoglycemia, hyperbilirubinemia, preeclampsia, etc. GDM patients and their fetuses have significantly increased risks of developing diabetes, obesity, and metabolic syndrome in the future, which not only increases the liv...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N30/89G01N30/72G01N30/88G01N30/06G01N30/08G01N30/14
CPCG01N33/6893G01N33/577G01N30/89G01N30/72G01N30/88G01N30/06G01N30/08G01N30/14G01N2800/042G01N2800/368G01N2570/00G01N2560/00G01N2030/8831G01N2030/062G01N2030/146G01N2030/067
Inventor 不公告发明人
Owner 安肽和(杭州)医疗科技有限公司
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