CIRCULATING SERUM MICRO-RNA AS NON-INVASIVE DIAGNOSTIC BIOMARKERS OF ENDOMETRIOSIS

Total RNA was extracted from 400 ul of serum with the miRNeasy Serum/Plasma advanced Kit (Qiagen) and quantified using the Qubit microRNA assay kit (Thermo Fisher Scientific). cDNA templates were prepared using the TaqMan Advanced miRNA cDNA Synthesis Kit (Thermo Fisher Scientific), starting from 10 ng of RNA. RT-qPCR carried out on a QuantStudio 12K Flex (Applied Biosystems) using the TaqMan OpenArray miRNA panel. This study was conducted on serum samples collected prior to surgery from 67 patients with endometriosis and 60 control women with benign gynecological conditions not linked to endometriosis, as confirmed by histological examination.

本研究采用miRNeasy血清/血浆高级提取试剂盒（miRNeasy Serum/Plasma advanced Kit，Qiagen）从400微升血清中提取总RNA，并通过Qubit microRNA检测试剂盒（Qubit microRNA assay kit，Thermo Fisher Scientific）完成定量。以10纳克总RNA为起始模板，使用TaqMan Advanced miRNA cDNA合成试剂盒（TaqMan Advanced miRNA cDNA Synthesis Kit，Thermo Fisher Scientific）制备cDNA模板。随后，借助TaqMan OpenArray miRNA芯片面板，在QuantStudio 12K Flex实时荧光定量PCR系统（Applied Biosystems）上开展逆转录实时荧光定量PCR（RT-qPCR）实验。本研究的血清样本均采集自术前受试者，涵盖67例子宫内膜异位症患者与60例经组织病理学检查确认无子宫内膜异位症相关良性妇科疾病的女性对照者。