OBJECTIVE To identify novel candidate diagnostic microRNA (miRNA) markers of endometriosis by means of an unbiased search with confirmation by means of targeted polymerase chain reaction (PCR). DESIGN Retrospective cohort. SETTING University teaching hospitals. PATIENT(S) Women with endometriosis and control women, confirmed with the use of laparoscopy. INTERVENTIONS(S) Diagnostic laparoscopy and blood sample. MAIN OUTCOME MEASURE(S) Next-generation sequencing (NGS) and quantitative real-time PCR (qRT-PCR). RESULT(S) Candidate miRNAs differentially expressed in women with endometriosis compared with control women were identified by means of NGS and selected for qRT-PCR. Plasma samples from another cohort of women with surgically confirmed endometriosis (n = 53) and disease-free control women (n = 53) were checked for hemolysis using spectrophotometry and the ratio of miR-23a and miR-451 by means of qRT-PCR. MicroRNA signatures were quantified by means of qRT-PCR in hemolysis-free plasma samples of case subjects (n = 25) and control subjects (n = 28) with the use of miRcury LNA miRNA. Circulating levels of eight miRNAs (miR-199a-3p, miR-143-3p, miR-340-5p, let-7b-5p, miR-21-5p, miR-17-5p, miR-20a-5p, and miR-103a-3p) were significantly lower in case subjects compared to control subjects. The sensitivity and specificity for individual miRNAs ranged from 0.36 to 1.00 and from 0.43 to 1.00, respectively, but when combined produced sensitivity and specificity of 0.92 and 0.86 with positive (PPV) and (NPV) predictive values of 0.85 and 0.92, respectively. However, combination of five miRNAs (miR-17-5p, miR-20a-5p, miR-199a-3p, miR-143-3p, and let-7b-5p) produced sensitivity and specificity of 0.96 and 0.79 with PPV and NPV of 0.80 and 0.96, respectively. CONCLUSION(S) We conclude that a panel of candidate miRNAs was comparable to laparoscopy in distinguishing between women with endometriosis and control women.

中文翻译：

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使用新一代测序和定量实时聚合酶链反应鉴定子宫内膜异位症的候选 microRNA 标记物

目的 通过无偏搜索和靶向聚合酶链反应 (PCR) 确认，鉴定子宫内膜异位症的新型候选诊断 microRNA (miRNA) 标记物。设计回顾性队列。设置大学教学医院。PATIENT(S) 子宫内膜异位症女性和对照女性，经腹腔镜检查确诊。干预措施(S) 诊断性腹腔镜检查和血液样本。主要结果测量 下一代测序 (NGS) 和定量实时 PCR (qRT-PCR)。结果 与对照女性相比，在子宫内膜异位症女性中差异表达的候选 miRNA 通过 NGS 鉴定并选择用于 qRT-PCR。使用分光光度法和通过 qRT-PCR 检测 miR-23a 和 miR-451 的比例，对来自另一组经手术确诊的子宫内膜异位症（n = 53）和无病对照女性（n = 53）的血浆样本进行了溶血检查. 使用 miRcury LNA miRNA，通过 qRT-PCR 在病例受试者 (n = 25) 和对照受试者 (n = 28) 的无溶血血浆样本中量化 MicroRNA 特征。8 种 miRNA（miR-199a-3p、miR-143-3p、miR-340-5p、let-7b-5p、miR-21-5p、miR-17-5p、miR-20a-5p 和 miR）的循环水平-103a-3p) 在病例受试者中显着低于对照受试者。单个 miRNA 的敏感性和特异性范围分别为 0.36 到 1.00 和 0.43 到 1.00，但当结合时产生的敏感性和特异性分别为 0.92 和 0。86 的阳性 (PPV) 和 (NPV) 预测值分别为 0.85 和 0.92。然而，五种 miRNA（miR-17-5p、miR-20a-5p、miR-199a-3p、miR-143-3p 和 let-7b-5p）的组合对 PPV 和 NPV 产生了 0.96 和 0.79 的敏感性和特异性分别为 0.80 和 0.96。结论（S） 我们得出结论，一组候选 miRNA 与腹腔镜检查在区分子宫内膜异位症女性和对照女性方面具有可比性。