BACKGROUND Cell-free mitochondrial DNA (cf-mtDNA) in body fluids has attracted much attention for the purpose of monitoring disease because of the clinical advantages. This study investigated whether the cf-mtDNA content in human follicular fluid samples was associated with oocyte and embryo developmental competence. METHODS We collected 225 individual follicular fluid samples from 92 patients undergoing conventional in vitro fertilization (n = 53) or intracytoplasmic sperm injection (n = 39). cf-mtDNA and cell-free nuclear DNA (cf-nDNA) were measured using real-time quantitative PCR for the ND1 and β-globin genes. Multivariate logistic regression and linear regression were used to analyze data. RESULTS The relative cf-mtDNA content (cf-ND1/cf-β-globin ratio) in follicular fluid was significantly lower in the group showing blastocyst development than in the non-blastocyst group (P = 0.030). Additionally, the relative cf-mtDNA content was significantly and positively correlated with the age of the female patient (P = 0.009), while the relative cf-mtDNA content for older women (≥38 years old) with anti-Müllerian hormone (AMH) ≤1.1 ng/ml was significantly higher than in those with AMH > 1.1 ng/ml (P <0.05). The cf-nDNA content was significantly positively correlated with the antral follicle count (P = 0.012), and significantly negatively correlated with both the number of days of stimulation and the total dose of gonadotropin administration (P = 0.039 and P = 0.015, respectively). Neither cf-mtDNA nor cf-nDNA levels in follicular fluid were associated with oocyte maturation, fertilization, or Day 3 embryo morphological scoring. CONCLUSIONS The relative cf-mtDNA content in human follicular fluid was negatively correlated with blastulation and positively correlated with the patient age, indicating that it is a promising bio-marker to evaluate oocyte developmental competence.

中文翻译：

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人卵泡液中的无细胞线粒体DNA：在接受辅助生殖技术的女性中，囊胚发育潜力的有前途的生物标志物。

背景技术体液中的无细胞线粒体DNA（cf-mtDNA）由于其临床优势而引起人们的广泛关注，以监测疾病。这项研究调查了人类卵泡液样本中的cf-mtDNA含量是否与卵母细胞和胚胎发育能力有关。方法我们收集了92例接受常规体外受精（n = 53）或胞浆内精子注射（n = 39）的患者的225个卵泡液样本。使用实时定量PCR对ND1和β-珠蛋白基因测量cf-mtDNA和无细胞核DNA（cf-nDNA）。使用多元逻辑回归和线性回归分析数据。结果显示胚泡发育的组卵泡液中cf-mtDNA的相对含量（cf-ND1 /cf-β-球蛋白比）显着低于非胚泡组（P = 0.030）。此外，相对cf-mtDNA含量与女性患者的年龄呈显着正相关（P = 0.009），而具有抗苗勒氏激素（AMH）的老年妇女（≥38岁）的相对cf-mtDNA含量≤1.1ng / ml显着高于AMH> 1.1 ng / ml的患者（P <0.05）。cf-nDNA含量与卵泡计数呈正相关（P = 0.012），与刺激天数和促性腺激素总剂量均呈显着负相关（分别为P = 0.039和P = 0.015） 。卵泡液中的cf-mtDNA和cf-nDNA含量均与卵母细胞成熟，受精或第3天胚胎形态学评分无关。结论人卵泡液中cf-mtDNA的相对含量与囊胚呈负相关，与患者年龄呈正相关，表明它是评估卵母细胞发育能力的有前途的生物标志物。