Diagnosis of pancreatic adenocarcinoma by polymerase chain reaction from pancreatic secretions

Multiple Mutation Analyses in Single Tumor Cells with Improved Whole Genome Amplification

“…Although detection of a Ki-ras mutation in codon 12 is generally critical due to the artificial introduction of Taq polymerase-introduced mutations, 38 this problem seems to be largely solved now, as we could demonstrate accurate mutation detection in codon 12 of the Ki-ras gene both in microdissected paraffin-embedded cells and microdissected disseminated tumor cells from bone marrow after immunostaining on cytospins. In contrast to the detection of tumor cells in blood or bone marrow by RT-PCR, [51][52][53][54][55] we have established a technique to characterize isolated disseminated tumor cells by multiple DNA analyses.…”

“…Detection of Ki-ras mutation in tumor tissue and in CK18-positive tumor cells from bone marrow of one pancreatic carcinoma patient and one rectal carcinoma patient was performed by enrichment PCR according to Trü mper et al 38 with minor modifications. Corresponding normal or CK18-negative cell DNA was used as a negative control; the CAPAN-1 cell line (American Type Culture Collection) served as a positive control.…”

“…Disseminated tumor cells were detected immunocytochemically by CK18 staining, picked from immunospots as single or few cells (n cell ϭ 3-5), and used for detection of Ki-ras mutation by RFLP-PCR according to Trü mper et al 38 The mutation analysis was also performed with microdissected tumor cells (n cell ϳ 200) from the formalin-fixed, paraffin-embedded primary tumors of these patients.…”

Multiplex PCR/LDR for detection of K-ras mutations in primary colon tumors

“…Samples were PCR ampli®ed and subjected to a multiplex ligase detection reaction (LDR), using the 26 primer set and the K294R mutant ligase enzyme as described in Materials and methods. The respective LDR product corresponds to the presence of the speci®c K-ras mutation listed at the top of the ®gure Discussion Detection of tumor-speci®c mutations in blood, urine, pancreatic secretions, and stool of cancer patients raises the exciting possibility of improved cancer screening, diagnosis, and staging using non-invasive molecular tests (Berthelemy et al, 1995;Caldas et al, 1994;Hasegawa et al, 1995;Mao et al, 1996;Nawroz et al, 1996;Nollau et al, 1996;Sidransky et al, 1992;Tada et al, 1993;Trumper et al, 1994;Wu et al, 1994). Mutations can serve as clonal markers of shed or disseminated cancer cells and are excellent targets for diagnostic testing Brennan et al, 1995;Hayashi et al, 1994Hayashi et al, , 1995Mao et al, 1994).…”

Methylation status of p14ARF and p16INK4a as detected in pancreatic secretions

“…Several studies have demonstrated the possibility to detect these mutations in pancreatic juice specimens or duodenal fluid (Tada et al, 1993;Trümper et al, 1994;Berthelemy et al, 1995;Van Laethem et al, 1995). However, further work revealed that k-ras mutations are also associated with benign or precancerous ductal lesions of unclear prognostic significance, thus limiting the clinical significance of detected k-ras mutations (Yanagisawa et al, 1993;Tada et al, 1996;Furuya et al, 1997).…”