title: Routine detection of the replication error phenotype in clinical tumor specimens using fluorescence-SSCP.
abstract: Almost all tumors from patients with hereditary non-polyposis colon carcinoma and approximately 10%-15% of sporadic colon and gastric carcinomas contain widespread deletions within mono- and dinucleotide repeat sequences in their DNA. This is referred to as the replication error (RER+) phenotype and, in the case of colon carcinoma, is often associated with an improved tumor prognosis and possibly also with response to chemotherapy. The RER+ status of tumors is usually determined by examining several dinucleotide and mononucleotide repeats for size variations when compared with the matching normal DNA. Alternately, the identification of deletions within BAT-26, a quasi-monomorphic polyadenine tract within the hMSH2 gene, has been shown to establish the RER+ status of tumors with greater than 99% accuracy and without the need for normal DNA. Here, we use fluorescent PCR in combination with single strand conformation polymorphism analysis to detect deletions in BAT-26. This technique provides a sensitive, rapid, reproducible and inexpensive assay suitable for the routine identification of RER+ status in clinical tumor specimens.
authors.affiliation: Department of Surgery, University of Western Australia, Nedlands, Australia. bjiac@cyllene.uwa.edu.au
authors.forename: B
authors.initials: B
authors.lastname: Iacopetta
authors.forename: F
authors.initials: F
authors.lastname: Grieu
mesh_terms: D004260:DNA Repair / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis; D004268:DNA-Binding Proteins; D005453:Fluorescence; D006801:Humans; D018895:Microsatellite Repeats; D051718:MutS Homolog 2 Protein; D009369:Neoplasms / Q000235:genetics*; D010641:Phenotype; D018807:Polymorphism, Single-Stranded Conformational; D011518:Proto-Oncogene Proteins / Q000235:genetics
chemical_list: D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; C497172:MSH2 protein, human; D051718:MutS Homolog 2 Protein