IntroductionEctopic pregnancy (EP) poses significant health risks, particularly in developing nations, necessitating improved diagnostic methods. This study aimed to explore potential biomarkers for EP diagnosis.MethodsA case–control study was conducted at the Sri Ramachandra Institute of Higher Education and Research, Chennai, Tamil Nadu. It included 140 EP cases and 140 pregnant controls, aged 19–38 years, attending routine visits. Serum samples were analysed for beta-human chorionic gonadotropin (β-hCG), progesterone, soluble fms-like tyrosine kinase-1 (sFLT-1) and eight microRNAs (miRs).ResultsDifferential expression of biomarkers was observed in EP cases. Four miRs (hsa-miR-141, hsa-miR-218, hsa-miR-519d and hsa-miR-873) were downregulated, and four miRs (hsa-miR-223, hsa-miR-517a, hsa-miR-523 and hsa-miR-323-3p) were upregulated. Statistically significant expression fold changes were noted (p<0.05), except for hsa-miR-141 and hsa-miR-218. miR-519d exhibited promising diagnostic potential with the highest specificity (97.1%) and a sensitivity of 47.1%. sFLT-1, as an individual marker, demonstrated a sensitivity of 98.6% and a specificity of 90%. The combination of sFLT-1 and miR-519d significantly enhanced the sensitivity to 100% with a specificity of 87.1%.ConclusionsThe combination of miR-519d and sFLT-1 emerges as a promising diagnostic tool for EP, offering a sensitivity of 100% and a specificity of 87.1%. These findings underscore the potential of biomarker-based approaches in improving EP diagnosis, especially in resource-limited settings. Further validation and clinical implementation studies are warranted to corroborate these findings and enhance EP management strategies.