Serum/blood plasma polypeptide marker related to gestational diabetes mellitus auxiliary early diagnosis and application thereof

A technology for early diagnosis and diabetes, applied in the field of proteomics, to quickly and accurately grasp the patient's condition, reduce adverse pregnancy outcomes, and reduce morbidity

Active Publication Date: 2018-12-07
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, there are currently no reports of relatively stable biomarkers for GDM-assisted early diagnosis. If GDM-specific or abnormally expressed serum/plasma peptide

Method used

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  • Serum/blood plasma polypeptide marker related to gestational diabetes mellitus auxiliary early diagnosis and application thereof
  • Serum/blood plasma polypeptide marker related to gestational diabetes mellitus auxiliary early diagnosis and application thereof
  • Serum/blood plasma polypeptide marker related to gestational diabetes mellitus auxiliary early diagnosis and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the collection of sample and the arrangement of sample data

[0026] The inventor collected a large number of peripheral blood samples from pregnant women at 14-18 weeks of pregnancy from Nanjing Maternal and Child Health Hospital from April to August 2015 (the samples used for research were collected at the same period, and the sampling, packaging, and storage conditions were uniform). By sorting out the sample data, the inventor selected 200 samples that meet the following criteria as experimental samples:

[0027] 1. Healthy pregnant women at 14-18 weeks of gestation at the time of blood collection

[0028] 2. Pregnant women who were diagnosed as GDM by OGTT during GDM screening at 24-28 weeks of gestation were defined as cases

[0029] 3. The above-mentioned research subjects did not develop GDM during GDM screening at 24-28 weeks of gestation. Healthy pregnant women who matched the age, BMI, and gestational weeks of the cases were defined as controls...

Embodiment 2

[0030] Embodiment 2, sample preparation, proteomics (peptidomics) screening and bioinformatics analysis

[0031] The above 100 eligible GDM cases and 100 healthy controls were subjected to ultrafiltration, TMT, LC-MS / MS and other experiments. The specific steps are as follows:

[0032] Collection preparation and ultrafiltration of samples in serum / plasma

[0033] 1. Centrifuge the sample at 3,000 rpm for 15 minutes at 4°C, and store the upper layer at -20°C for further analysis. Usually 200-250ul serum or plasma can be obtained.

[0034] 2. Centrifuge the sample at 12,000 revolutions per minute at 4°C for 15 minutes, add acrylcyanide to the centrifuged supernatant, then vortex briefly, place it at room temperature for 20 minutes, and then use it according to the instructions, using 10kDa MWCO Peptides below 10 kDa were obtained by filter (Millipore, USA) and lyophilized.

[0035] 3. The protein concentration was measured by using BCA (Pierce, Rockford, IL, USA).

[0036] 4...

Embodiment 3

[0060] Embodiment 3, random verification

[0061] According to the above results, 7 differential polypeptides that were significantly highly expressed in the GDM outcome group or only expressed in the GDM outcome group were randomly selected as verification targets. In our early established serum bank at 14-18 weeks of pregnancy, the normal outcome group was selected. and 30 serum samples from the GDM outcome group (the two groups matched each other in terms of age, BMI, and gestational age), further verifying the results of our preliminary screening, and the study confirmed that 4 of the 7 peptides (Q6PFW1, P35658, P32004, and Q9Y4H2 ) expression patterns are basically consistent with the results of primary screening (Table 2), and there are significant differences in the expression between the two groups, suggesting that they are associated with the onset of gestational diabetes mellitus, and they may become molecular markers for early prediction and diagnosis of GDM ( figu...

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Abstract

The invention discloses a serum/blood plasma polypeptide marker related to gestational diabetes mellitus auxiliary early diagnosis and application thereof. The serum/blood plasma polypeptide marker ischaracterized in that the marker is selected from arbitrary one or more polypeptides of Q6PFW1, P35658, P32004 and Q9Y4H2. The serum/blood plasma polypeptide marker serving as a detection target is applied to preparation of gestational diabetes mellitus auxiliary early diagnosis reagents. The expression quantity of the marker for detecting pregnancy serum/blood plasma can be used in GDM auxiliaryearly diagnosis. Support is provided to clinical doctors for accurately predicting the GDM pathogenesis outcome of pregnancy and timely adopting personal prevention and treatment schemes, so that theoccurrence rate of GDM can be furthest reduced, and adverse pregnancy outcomes can be reduced.

Description

technical field [0001] The invention belongs to the field of proteomics, and relates to serum / plasma polypeptide markers and applications thereof related to auxiliary early diagnosis of gestational diabetes. Background technique [0002] Gestational diabetes mellitus (GDM) refers to varying degrees of abnormal glucose tolerance that occurs or is discovered for the first time during pregnancy. If it occurs in the first trimester of pregnancy, the possibility of abnormal glucose tolerance already existing before pregnancy cannot be ruled out. GDM is one of the most common complications during pregnancy. About 3%-8% of pregnant women develop GDM during pregnancy. It is particularly noteworthy that the incidence of GDM continues to rise with the increase in the obesity rate of women of childbearing age. [0003] GDM can bring a series of complications to the mother and fetus, such as increasing the incidence of spontaneous abortion, pregnancy-induced hypertension, polyhydramnios...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/689G01N2800/368
Inventor 石中华沈嵘丁虹娟蒋涛尹玲凤赵纯单春剑
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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