title: Shotgun proteomics identifies serum fibronectin as a candidate diagnostic biomarker for inclusion in future multiplex tests for ectopic pregnancy.
abstract: Ectopic pregnancy (EP) is difficult to diagnose early and accurately. Women often present at emergency departments in early pregnancy with a 'pregnancy of unknown location' (PUL), and diagnosis and exclusion of EP is challenging due to a lack of reliable biomarkers. The objective of this study was to identify novel diagnostic biomarkers for EP. Shotgun proteomics, incorporating combinatorial-ligand library pre-fractionation, was used to interrogate pooled sera (n = 40) from women undergoing surgery for EP, termination of viable intrauterine pregnancy and management of non-viable intrauterine pregnancy. Western blot was used to validate results in individual sera. ELISAs were developed to interrogate sera from women with PUL (n = 120). Sera were collected at time of first symptomatic presentation and categorized according to pregnancy outcome. The main outcome measures were differences between groups and area under the receiver operating curve (ROC). Proteomics identified six biomarker candidates. Western blot detected significant differences in levels of two of these candidates. ELISA of sera from second cohort revealed that these differences were only significant for one of these candidates, fibronectin. ROC analysis of ability of fibronectin to discriminate EP from other pregnancy outcomes suggested that fibronectin has diagnostic potential (ROC 0.6439; 95% CI 0.5090 to 0.7788; P>0.05), becoming significant when 'ambiguous' medically managed PUL excluded from analysis (ROC 0.6538; 95% CI 0.5158 to 0.7918; P<0.05). Fibronectin may make a useful adjunct to future multiplex EP diagnostic tests.
authors.affiliation: Medical Research Council Centre for Reproductive Health, The University of Edinburgh, Queen's Medical Research Institute, Edinburgh, United Kingdom.
authors.forename: Jeremy K
authors.initials: JK
authors.lastname: Brown
authors.forename: Katarina B
authors.initials: KB
authors.lastname: Lauer
authors.forename: Emily L
authors.initials: EL
authors.lastname: Ironmonger
authors.forename: Neil F
authors.initials: NF
authors.lastname: Inglis
authors.forename: Tom H
authors.initials: TH
authors.lastname: Bourne
authors.forename: Hilary O D
authors.initials: HO
authors.lastname: Critchley
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Horne
mesh_terms: D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000097:blood; D015153:Blotting, Western; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005353:Fibronectins / Q000097:blood*; D006801:Humans; D008875:Middle Aged; D010449:Peptide Mapping; D011247:Pregnancy; D011256:Pregnancy Outcome; D011258:Pregnancy Tests / Q000379:methods*; D011271:Pregnancy, Ectopic / Q000097:blood* / Q000175:diagnosis* / Q000378:metabolism / Q000601:surgery; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D055815:Young Adult
chemical_list: D015415:Biomarkers; D005353:Fibronectins