Novel Approach to Fecal Occult Blood Testing by Assay of Erythrocyte-Specific microRNA Markers

Chung Wah Wu, Xiaoming Cao, Calise K. Berger, Patrick H. Foote, Douglas W. Mahoney, Julie A. Simonson, Bradley W. Anderson, Tracy C. Yab, William R. Taylor, Lisa A. Boardman, John B. Kisiel, David A. Ahlquist

Research output: Contribution to journalArticlepeer-review

14 Scopus citations


Background: Fecal occult blood testing (FOBT) has historically relied on methods to detect hemoglobin with no fundamental innovations in decades. Aim: To examine microRNA (miRNA) as a new marker class for FOBT. Methods: Candidate miRNA markers were identified by small RNA sequencing of human whole blood compared to colorectal epithelia. Markers were tested in human blood cell subsets and blood from non-human species. We assessed assay linearity in blood spiking and marker stability in stool over incubation experiments. Levels of candidate erythrocyte markers were explored in stools from colorectal cancer (CRC) cases and controls. Results: Based on small RNA sequencing and validation RT-qPCR, expression level of each of the top blood-enriched markers (hsa-miR-144-3p, 144-5p, 451a, 486-5p, 363-3p, 20b-5p) could perfectly discriminate blood from colorectal epithelia. All six markers arose from and showed specificity to human erythrocytes. Marker levels increased linearly with erythrocyte concentration in saline or stool and demonstrated a broader dynamic range than did immunochemical test for hemoglobin. Degradation of markers occurred in stool but was reduced with preservative buffers. Erythrocyte marker candidates for stool testing were selected in an exploratory set of stools (20 CRC, 40 normal). Candidates were then further tested in a feasibility set (29 CRC, 31 advanced adenoma, and 115 normal); a miRNA panel (hsa-miR-451a, 144-5p, and 200b-3p as normalizer) yielded an AUC of 0.89 (95% CI 0.82–0.95, P < .0001) for CRC. Conclusions: A novel miRNA-based approach accurately quantifies fecal blood levels over a broad, clinically relevant range.

Original languageEnglish (US)
Pages (from-to)1985-1994
Number of pages10
JournalDigestive diseases and sciences
Issue number8
StatePublished - Aug 1 2017


  • Biomarkers
  • Colorectal neoplasms
  • Erythrocytes
  • Occult blood
  • microRNAs

ASJC Scopus subject areas

  • Physiology
  • Gastroenterology


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