title: PCR-detection of tumor-derived p53 DNA in cerebrospinal fluid.
abstract: The sensitivity of polymerase chain reaction (PCR)-based methods for the detection of DNA offers opportunities for tumor diagnosis from the small amounts of tumor-derived DNA released into body fluids. Tumor-derived DNA can be distinguished from DNA derived from non-neoplastic cells by the presence of tumor specific genomic alterations, such as mutations in the p53 gene. This case report describes the use of allele-specific PCR (A-PCR) to detect a C-->T transition in p53 codon 273 in DNA extracted from the cerebrospinal fluid (CSF) of a patient whose glioblastoma contained the same mutation. The results of this study were confirmed by a second independent A-PCR reaction that detected the corresponding G-->A transition on the opposite strand. The specificity of the A-PCR protocol was demonstrated by negative controls, including pooled human placental DNA and the patient's non-tumor DNA, and by the use of A-PCR primers to detect all four possible bases at the site of the mutation. The methodology used in this study is suitable for use as a diagnostic clinical test. Because about half of all human tumors contain p53 mutations, PCR examination of CSF for the presence of mutant p53 sequences may be useful in the diagnosis of recurrent or metastatic tumors. Patients with known carcinoma of the breast or lung might be particularly benefited by this test.
authors.affiliation: Department of Pathology, Dartmouth-Hitchcock Medical Center, Lebanon, New Hampshire 03736, USA.
authors.forename: C H
authors.initials: CH
authors.lastname: Rhodes
authors.forename: C
authors.initials: C
authors.lastname: Honsinger
authors.forename: G D
authors.initials: GD
authors.lastname: Sorenson
mesh_terms: D000293:Adolescent; D001483:Base Sequence; D001932:Brain Neoplasms / Q000134:cerebrospinal fluid* / Q000235:genetics; D004273:DNA, Neoplasm / Q000134:cerebrospinal fluid*; D005091:Exons; D005909:Glioblastoma / Q000134:cerebrospinal fluid* / Q000235:genetics; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm; D015345:Oligonucleotide Probes; D016159:Tumor Suppressor Protein p53