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Reference gene combination for gene expression analysis of polycystic ovarian syndrome and application thereof

A polycystic ovary, internal reference gene technology, applied in the fields of molecular biology and disease gene research and diagnosis, can solve problems such as underestimation or overestimation of gene expression

Active Publication Date: 2017-10-17
SDIVF R&D CENT LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using inappropriate housekeeping genes may lead to data distortion and underestimation or overestimation of gene expression (Dheda, et al., 2005; Murphy and Bustin, 2009; Nestorov, et al., 2013)

Method used

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  • Reference gene combination for gene expression analysis of polycystic ovarian syndrome and application thereof
  • Reference gene combination for gene expression analysis of polycystic ovarian syndrome and application thereof
  • Reference gene combination for gene expression analysis of polycystic ovarian syndrome and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] call for researchers

[0025] From January 2015 to June 2016, this study involved 89 Chinese Han women, aged 29.12±3.01 years, from the Affiliated Hospital of Shandong University. They included 44 women with polycystic ovary syndrome (PCOS) and 45 ovulatory women undergoing tubal and / or male infertility treatment. All patients or family members provided written consent. This study was approved by the ethics committee.

Embodiment 2

[0027] Clinical and Biochemical Testing

[0028] Polycystic ovary syndrome (PCOS) was diagnosed according to the 2003 Rotterdam criteria (Chang, et al., 2004; Fauser, et al., 2004), including any two of the following three clinical features: 1. Ovulation / anovulation; 2. Clinical and / or biochemical hyperandrogenism; 3. Ultrasound examination of polycystic ovary, and exclusion of other pathophysiological conditions associated with hyperandrogenism, including congenital adrenal hyperplasia, Cushing's syndrome, and androgen-secreting tumors.

[0029] All relevant research subjects measured their body parameters, such as waist circumference, hip circumference, height and weight, when they first visited the outpatient clinic. Body mass index (BMI) is calculated as weight (kg) divided by height squared (m 2 ), or BMI=kg / m 2 . Venous blood samples were collected between 8:00 and 10:00 am after a 12-hour fast. All blood samples from women with PCOS were obtained during the early fo...

Embodiment 3

[0037] Granulosa Cell Isolation

[0038] Granulosa cells were isolated from 44 women with polycystic ovary syndrome (PCOS) and a control group of 45 normal ovulatory women undergoing tubal and / or male infertility treatment. Volunteers with a history of other gynecological or medical diseases were excluded. All women were injected with a gonadotropin-releasing hormone (GnRH) agonist starting in the midluteal phase, and follicle size was monitored by ultrasonography and serum estradiol measurements. When three or more follicles with an average diameter of R1.8 cm were detected, 8000-10000 IU of human chorionic gonadotropin (Profasi, Serono) was administered 36 hours before ultrasound-guided oocyte retrieval. After anesthesia, oocytes were retrieved via a 17-gauge double-lumen puncture needle (K-OPS-WOOD-1235, Cook Australia). Granulosa cells (GCs) were collected around the oocytes, removed by suction with a Pasteur pipette, and washed twice with Dulbecco's modified Eagle mediu...

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Abstract

The invention provides application of HPRT1 (hypoxanthine-guanine phosphoribosyltransferase 1), RPLP0 (ribosomal protein, large, P0) and HMBS (hydroxymethylbilane synthase) in the preparation as a reference gene in the preparation of kits or devices for studying or detecting polycystic ovarian syndrome. The invention also provides application of the HPRT1, RPLP0 and HMBS combination as kits or devices for studying or detecting hydroxymethylbilane synthase, such as real-time fluorescence quantitative PCR kits or gene chips.

Description

technical field [0001] The invention relates to the fields of molecular biology and disease gene research and diagnosis. Specifically, the present invention relates to an internal reference gene combination used in the study of polycystic ovary syndrome (PCOS) gene expression analysis and an internal reference gene combination for detecting polycystic ovary syndrome. Background technique [0002] Polycystic ovary syndrome (PCOS), whose symptoms include ovulatory dysfunction, polycystic ovaries, and androgen excess, is a common gynecological endocrine disorder and a major cause of female reproductive failure (Kovacs and Wood, 2001; Wild, et al. , 2000). According to different diagnostic criteria, its prevalence is estimated to be 6% to 18% (Azziz, et al., 2004; Christian, et al., 2003; March, et al., 2010). PCOS patients are prone to multiple metabolic disorders including insulin resistance, type 2 diabetes mellitus (T2DM), obesity, dyslipidemia, chronic low-level inflammat...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/158C12Q2600/166
Inventor 陈子江马金龙刘洪彬吕跃赵世刚熊志强路钢
Owner SDIVF R&D CENT LTD
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