title: Stool DNA testing for the detection of pancreatic cancer: assessment of methylation marker candidates.
abstract: BACKGROUND
  Pancreatic cancer (PanC) presents at late stage with high mortality. Effective early detection methods are needed. Aberrantly methylated genes are unexplored as markers for noninvasive detection by stool testing. The authors aimed to select discriminant methylated genes and to assess accuracy of these and mutant KRAS in stool to detect PanC.
  
  
  METHODS
  Nine target genes were assayed by real-time methylation-specific polymerase chain reaction (MSP) in bisulfite-treated DNA from microdissected frozen specimens of 24 PanC cases and 30 normal colon controls. Archived stools from 58 PanC cases and 65 controls matched on sex, age, and smoking were analyzed. Target genes from fecal supernatants were enriched by hybrid capture, bisulfite-treated, and assayed by MSP. KRAS mutations were assayed using the QuARTS technique.
  
  
  RESULTS
  Areas under the receiver operating characteristics curves (AUCs) for tissue BMP3, NDRG4, EYA4, UCHL1, MDFI, Vimentin, CNTNAP2, SFRP2, and TFPI2 were 0.90, 0.79, 0.78, 0.78, 0.77, 0.77, 0.69, 0.67, and 0.66, respectively. The top 4 markers and mutant KRAS were evaluated in stool. BMP3 was the most discriminant methylation marker in stool. At 90% specificity, methylated BMP3 alone detected 51% of PanCs, mutant KRAS detected 50%, and combination detected 67%. AUCs for methylated BMP3, mutant KRAS, and combination in stool were 0.73, 0.75, and 0.85, respectively.
  
  
  CONCLUSIONS
  This study demonstrates that stool assay of a methylated gene marker can detect PanC. Among candidate methylated markers discriminant in tissue, BMP3 alone performed well in stool. Combining methylated BMP3 and mutant KRAS increased stool detection over either marker alone.
authors.affiliation: Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA.
authors.forename: John B
authors.initials: JB
authors.lastname: Kisiel
authors.forename: Tracy C
authors.initials: TC
authors.lastname: Yab
authors.forename: William R
authors.initials: WR
authors.lastname: Taylor
authors.forename: Suresh T
authors.initials: ST
authors.lastname: Chari
authors.forename: Gloria M
authors.initials: GM
authors.lastname: Petersen
authors.forename: Douglas W
authors.initials: DW
authors.lastname: Mahoney
authors.forename: David A
authors.initials: DA
authors.lastname: Ahlquist
mesh_terms: D019540:Area Under Curve; D015415:Biomarkers; D055398:Bone Morphogenetic Protein 3 / Q000235:genetics; D018395:CA-19-9 Antigen / Q000097:blood; D016022:Case-Control Studies; D004247:DNA / Q000032:analysis*; D019175:DNA Methylation; D005243:Feces / Q000737:chemistry*; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D010190:Pancreatic Neoplasms / Q000175:diagnosis* / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins / Q000235:genetics
chemical_list: C525757:BMP3 protein, human; D015415:Biomarkers; D055398:Bone Morphogenetic Protein 3; D018395:CA-19-9 Antigen; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D004247:DNA; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins
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