title: Clinical performance of an automated stool DNA assay for detection of colorectal neoplasia.
abstract: OBJECTIVE
  Colorectal cancer (CRC) and advanced precancers can be detected noninvasively by analyses of exfoliated DNA markers and hemoglobin in stool. Practical and cost-effective application of a stool DNA-based (sDNA) test for general CRC screening requires high levels of accuracy and high-capacity throughput. We optimized an automated sDNA assay and evaluated its clinical performance.
  
  
  METHODS
  In a blinded, multicenter, case-control study, we collected stools from 459 asymptomatic patients before screening or surveillance colonoscopies and from 544 referred patients. Cases included CRC (n = 93), advanced adenoma (AA) (n = 84), or sessile serrated adenoma ≥1 cm (SSA) (n = 30); controls included nonadvanced polyps (n = 155) or no colonic lesions (n = 641). Samples were analyzed by using an automated multi-target sDNA assay to measure β-actin (a marker of total human DNA), mutant KRAS, aberrantly methylated BMP3 and NDRG4, and fecal hemoglobin. Data were analyzed by a logistic algorithm to categorize patients as positive or negative for advanced colorectal neoplasia (CRC, advanced adenoma, and/or SSA ≥1 cm).
  
  
  RESULTS
  At 90% specificity, sDNA analysis identified individuals with CRC with 98% sensitivity. Its sensitivity for stage I cancer was 95%, for stage II cancer it was 100%, for stage III cancer it was 96%, for stage IV cancer it was 100%, and for stages I-III cancers it was 97% (nonsignificant P value). Its sensitivity for advanced precancers (AA and SSA) ≥1 cm was 57%, for >2 cm it was 73%, and for >3 cm it was 83%. The assay detected AA with high-grade dysplasia with 83% sensitivity.
  
  
  CONCLUSIONS
  We developed an automated, multi-target sDNA assay that detects CRC and premalignant lesions with levels of accuracy previously demonstrated with a manual process. This automated high-throughput system could be a widely accessible noninvasive approach to general CRC screening.
authors: Exact Sciences Corporation, Madison, Wisconsin.
authors: Graham P
authors: GP
authors: Lidgard
authors: Michael J
authors: MJ
authors: Domanico
authors: Janelle J
authors: JJ
authors: Bruinsma
authors: James
authors: J
authors: Light
authors: Zubin D
authors: ZD
authors: Gagrat
authors: Rebecca L
authors: RL
authors: Oldham-Haltom
authors: Keith D
authors: KD
authors: Fourrier
authors: Hatim
authors: H
authors: Allawi
authors: Tracy C
authors: TC
authors: Yab
authors: William R
authors: WR
authors: Taylor
authors: Julie A
authors: JA
authors: Simonson
authors: Mary
authors: M
authors: Devens
authors: Russell I
authors: RI
authors: Heigh
authors: David A
authors: DA
authors: Ahlquist
authors: Barry M
authors: BM
authors: Berger
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D057205:Automation, Laboratory / Q000379:methods*; D014408:Biomarkers, Tumor / Q000032:analysis*; D016022:Case-Control Studies; D015179:Colorectal Neoplasms / Q000175:diagnosis*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000737:chemistry*; D005260:Female; D006454:Hemoglobins / Q000032:analysis*; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D016037:Single-Blind Method
chemical_list: D014408:Biomarkers, Tumor; D006454:Hemoglobins; D004247:DNA
keywords: AA; BMP3; CRC; Colon Cancer; Early Detection; FIT; HGD; Hb; NDRG4; QuARTS; SSA; advanced adenoma; colorectal cancer; fecal immunochemical test; hemoglobin; high-grade dysplasia; quantitative allele-specific real-time target and signal amplification; sDNA; sessile serrated adenoma ≥1-cm size; stool DNA