title: Application of the fecal microRNA test to the residuum from the fecal occult blood test.
abstract: OBJECTIVE
  Though the fecal occult blood test is used for colorectal cancer screening worldwide, it does not have a particularly high sensitivity for detecting colorectal cancer. Here we investigated the applicability of the fecal microRNA test to fecal samples that had been used for a previous fecal occult blood test and stored under various conditions.
  
  
  METHODS
  Five colorectal cancer patients and five healthy volunteers were enrolled. Fecal samples were stored for 0-5 days at 4°C, room temperature or 37°C. Total RNA was extracted from the fecal occult blood test residuum and microRNA expression was analyzed by real-time reverse transcription polymerase chain reaction.
  
  
  RESULTS
  There were no remarkable differences either in colorectal cancer patients or in controls with regard to the concentration of RNA extracted from the fecal occult blood test residuum in any of the storage groups compared with the samples prepared on day 0 (Group 0). Ribosomal RNA stored at room temperature or 37°C degraded rapidly. In contrast, the ribosomal RNA stored at 4°C remained intact for at least 5 days. The microRNAs in samples stored at 4°C and room temperature were conserved; however, the microRNAs stored at 37°C were significantly degraded compared with Group 0 (P < 0.05). In the residuum stored at 4°C up to 5 days, the relative quantification of miR-106a normalized with miR-24 in colorectal cancer patients was significantly higher than those in healthy volunteers (P < 0.05). In contrast, the quantification of normalized miR-106a was remarkably low in samples stored at room temperature and 37°C.
  
  
  CONCLUSIONS
  Fecal microRNA of sufficient quality for reverse transcription polymerase chain reaction analysis was extracted from the fecal occult blood test residuum stored at 4°C for up to 5 days.
authors.affiliation: Department of Colorectal and Pelvic Surgery, National Cancer Center Hospital East, Kashiwa, Japan.
authors.forename: Nobuyoshi
authors.initials: N
authors.lastname: Yamazaki
authors.forename: Yoshikatsu
authors.initials: Y
authors.lastname: Koga
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Kakugawa
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Otake
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Hayashi
authors.forename: Norio
authors.initials: N
authors.lastname: Saito
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Matsumura
mesh_terms: D000328:Adult; D000368:Aged; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000517:prevention & control*; D005243:Feces / Q000737:chemistry*; D005260:Female; D006801:Humans; D008297:Male; D008403:Mass Screening / Q000379:methods*; D035683:MicroRNAs / Q000032:analysis* / Q000302:isolation & purification; D008875:Middle Aged; D009780:Occult Blood; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods
chemical_list: D035683:MicroRNAs
keywords: cancer screening; colorectal cancer; fecal miRNA; fecal occult blood test