Identification and quantification of preterm birth biomarkers in human cervicovaginal fluid by liquid chromatography/tandem mass spectrometry.

摘要: Spontaneous preterm birth (PTB) before 37 completed weeks of gestation resulting from labor (PTL) is a leading contributor perinatal morbidity and mortality. Early identification at-risk women by reliable screening tests could alleviate this health issue; however, conventional methods such as obstetric history clinical risk factors, uterine activity monitoring, biochemical markers, cervical sonography for at PTB have proven unsuccessful in lowering the rate PTB. Cervicovaginal fluid (CVF) might prove to be useful, readily available biological identifying diagnostic biomarkers. Human columnar epithelial endocervical-1 (End1) vaginal (Vk2) cell secretomes were employed generate stable isotope labeled proteome (SILAP) standard facilitate characterization relative quantification proteins present CVF. The SILAP was prepared using labeling amino acids culture (SILAC) End1 Vk2 through seven passages. secreted both lines combined characterized liquid-chromatography-tandem mass spectrometry (LC-MS/MS). In total, 1211 identified End1-Vk2 standard, with 236 being consistently each replicates analyzed. Individual found contain <0.5% endogenous unlabeled forms. Identified screened provide set 15 candidates that either previously been potential biomarkers or linked mechanistically Stable dilution LC-multiple reaction monitoring (MRM/MS) assays then developed conducting candidate human CVF samples term cases. Three significantly elevated cases (desmoplakin isoform 1, stratifin, thrombospondin 1 precursor), providing foundation further validation larger patient cohorts.