Detecting colorectal cancer in stool with the use of multiple genetic targets.
BACKGROUND
Colorectal cancer cells are shed into the stool, providing a potential means for the early detection of the disease using noninvasive approaches. Our goal was to develop reliable, specific molecular genetic tests for the detection of colorectal cancer in stool samples.


METHODS
Stool DNA was isolated from paired stools and primary tumor samples from 51 colorectal cancer patients. Three genetic targets-TP53, BAT26, and K-RAS-were used to detect tumor-associated mutations in the stool prior to or without regard to the molecular analyses of the paired tumors. TP53 gene mutations were detected with a mismatch-ligation assay that detects nine common p53 gene mutations. Deletions within the BAT26 locus were detected by a modified solid-phase minisequencing method. Mutations in codons 12 and 13 of K-RAS were detected with a digital polymerase chain reaction-based method.


RESULTS
TP53 gene mutations were detected in the tumor DNA of 30 patients, all of whom had the identical TP53 mutation in their stools. Tumors from three patients contained a noninherited deletion at the BAT26 locus, and the same alterations were identified in these patients' stool specimens. Nineteen of 50 tumors tested had a K-RAS mutation; identical mutations were detected in the paired stool DNA samples from eight patients. In no case was a mutation found in stool that was not also present in the primary tumor. Thus, the three genetic markers together detected 36 (71%) of 51 patients (95% confidence interval [CI] = 56% to 83%) with colorectal cancer and 36 (92%) of 39 patients (95% CI = 79% to 98%) whose tumors had an alteration.


CONCLUSIONS
We were able to detect the majority of colorectal cancers by analyzing stool DNA for just three genetic markers. Additional work is needed to determine the specificity of these genetic tests for detecting colorectal neoplasia in asymptomatic patients and to more precisely estimate the prevalence of the mutations and sensitivity of the assay.
Dong
S M
SM
Division of Head and Neck Cancer Research, Department of Otolaryngology-Head and Neck Surgery, The Johns Hopkins Medical School, The Johns Hopkins University, Baltimore, MD, USA.
Traverso
G
G
Johnson
C
C
Geng
L
L
Favis
R
R
Boynton
K
K
Hibi
K
K
Goodman
S N
SN
D'Allessio
M
M
Paty
P
P
Hamilton
S R
SR
Sidransky
D
D
Barany
F
F
Levin
B
B
Shuber
A
A
Kinzler
K W
KW
Vogelstein
B
B
Jen
J
J
D000368:Aged; D003110:Colonic Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D005243:Feces / Q000737:chemistry*; D016158:Genes, p53; D011905:Genes, ras; D005819:Genetic Markers; D018095:Germ-Line Mutation; D006801:Humans; D008875:Middle Aged; D009154:Mutation; D009367:Neoplasm Staging; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D017384:Sequence Deletion
D005819:Genetic Markers; D004247:DNA