TY - JOUR
T1 - Detecting low-abundance p16 and p53 mutations in pancreatic juice using a novel assay
T2 - Heteroduplex analysis of limiting dilution PCRs
AU - Bian, Yansong
AU - Matsubayashi, Hiroyuki
AU - Pin-Li, Chung
AU - Abe, Tadayoshi
AU - Canto, Marcia
AU - Murphy, Kathleen M.
AU - Goggins, Michael
N1 - Funding Information:
Supported by the SPORE grant in Gastro intestinal Cancer (CA62924), R01CA120432 from the National Cancer Institute, and the Michael Rolfe Foundation USA and has the lowest survival rate for any solid cancer (~2%).1,2This poor survival occurs in part because only ~15% of patients are diagnosed with pancreatic cancer while they have surgically resectable disease. Pancreatic cancer survival is better for patients with the smallest tumors.3-5 Such a poor survival is particularly of concern to patients with inherited susceptibility to the disease.6,7 Screening asymptomatic individuals with a significant risk of developing pancreatic cancer has demonstrated that preinvasive pancreatic neoplasms can be detected by endoscopic ultrasound in some individuals.8,9These results suggest that some form of clinical screening for high-risk individuals will eventually become an important part of their management.8,9
PY - 2006/10
Y1 - 2006/10
N2 - We have developed a simple, robust, highly-sensitive assay for identifying gene mutations in clinical samples. We applied this assay to detect p53 and p16 mutations in pancreatic juice obtained from patients undergoing evaluation and treatment of pancreatic disease. The assay strategy involves PCR amplifying DNA at limiting dilution (LD-PCR) followed by screening PCR products for mutations using temperature gradient capillary electrophoresis (TGCE). Compared to conventional TGCE, TGCE after LD-PCR significantly increased the number of detectable mutations in pancreatic duct juice. Using LD-PCR, mutations in p53 and/or p16 were found in the pancreatic juice of 12 of 20 individuals with pancreatic cancer compared to only 1 of 8 patients with chronic pancreatitis, 0 of 8 individuals without evidence of pancreatic disease (p < 0.02). We conclude that limiting dilution PCR is an effective strategy for improving the detection of mutations in clinical samples and when applied to pancreatic juice to detect mutations of p53 and/or p16, it can help distinguish patients with pancreatic cancer from those without evidence of pancreatic neoplasia.
AB - We have developed a simple, robust, highly-sensitive assay for identifying gene mutations in clinical samples. We applied this assay to detect p53 and p16 mutations in pancreatic juice obtained from patients undergoing evaluation and treatment of pancreatic disease. The assay strategy involves PCR amplifying DNA at limiting dilution (LD-PCR) followed by screening PCR products for mutations using temperature gradient capillary electrophoresis (TGCE). Compared to conventional TGCE, TGCE after LD-PCR significantly increased the number of detectable mutations in pancreatic duct juice. Using LD-PCR, mutations in p53 and/or p16 were found in the pancreatic juice of 12 of 20 individuals with pancreatic cancer compared to only 1 of 8 patients with chronic pancreatitis, 0 of 8 individuals without evidence of pancreatic disease (p < 0.02). We conclude that limiting dilution PCR is an effective strategy for improving the detection of mutations in clinical samples and when applied to pancreatic juice to detect mutations of p53 and/or p16, it can help distinguish patients with pancreatic cancer from those without evidence of pancreatic neoplasia.
KW - Limiting dilution
KW - Mutation
KW - Pancreatic adenocarcinoma
KW - Pancreatic juice
KW - Temperature gradient capillary electrophoresis
KW - p16
KW - p53
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U2 - 10.4161/cbt.5.10.3453
DO - 10.4161/cbt.5.10.3453
M3 - Article
C2 - 17106238
AN - SCOPUS:33751175168
SN - 1538-4047
VL - 5
SP - 1392
EP - 1399
JO - Cancer Biology and Therapy
JF - Cancer Biology and Therapy
IS - 10
ER -