ME-PCR for the identification of mutated K-ras in serum and bile of pancreatic cancer patients: an unsatisfactory technique for clinical applications

• Published in: Clinica chimica acta Vol. 302; no. 1; pp. 35 - 48
• Main Authors: Zambon, Carlo-Federico, Navaglia, Filippo, Basso, Daniela, Gallo, Nicoletta, Greco, Eliana, Piva, Maria Grazia, Fogar, Paola, Pasquali, Claudio, Pedrazzoli, Sergio, Plebani, Mario
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier B.V 01-12-2000; Elsevier
• Subjects: Adult; Aged; Aged, 80 and over; Bile; Bile - chemistry; Biological and medical sciences; Codon; DNA - analysis; DNA - blood; DNA Mutational Analysis - methods; Electrophoresis, Agar Gel; Female; Gastroenterology. Liver. Pancreas. Abdomen; Genes, ras; Humans; K-ras; Liver. Biliary tract. Portal circulation. Exocrine pancreas; Male; ME-PCR; Medical sciences; Middle Aged; Mutation; Pancreatic cancer; Pancreatic Neoplasms - genetics; Point Mutation; Polymerase Chain Reaction - methods; Serum; Tumor Cells, Cultured; Tumors; Serum; K-ras; ME-PCR; Pancreatic cancer; Bile; Human; Cell culture; Mutant enriched polymerase chain reaction; Biological marker; Malignant tumor; Blood; Polymerase chain reaction; Clinical biology; Digestive diseases; Diagnosis; Mutation; Molecular biology; Onc gene; Pancreas; Ras gene; Tumor cell; Pancreatic disease
• ISSN: 0009-8981; 1873-3492
• DOI: 10.1016/S0009-8981(00)00351-X

Our aim was to assess the clinical reliability of mutated
K-ras detection in serum or bile for the diagnosis of pancreatic cancer using ME-PCR. DNA was extracted from 1 ml serum obtained from 29 patients with pancreatic cancer and 12 control subjects. ME-PCR was optimized using a mixture of normal DNA added with different amounts of mutated DNA. The analysis of sera obtained from the 29 patients and of bile obtained from 11 pancreatic cancer patients demonstrated the presence of mutated
K-ras in two (6.9%) and four cases (36%). By contrast
K-ras was not amplifiable in any of the 12 serum samples obtained from healthy controls. In conclusion the DNA obtained from pancreatic cancer patients’ sera is suitable for
K-ras amplification and for the identification of codon 12 point mutations. However ME-PCR alone has an unsatisfactory sensitivity for the detection of pancreatic cancer using serum DNA as starting template.