Use of TP53 reference materials to validate mutations in clinical tissue specimens by single-strand conformational polymorphism analysis.
BACKGROUND
As genetic information moves from basic research laboratories in to the clinical testing environment, there is a critical need for reliable reference materials for the quality assurance of genetic tests. A panel of 12 plasmid clones containing wild-type or point mutations within exons 5-9 have been developed as reference materials for the detection of TP53 mutations.


OBJECTIVE
The goal of this study was to validate the reference materials in providing quality assurance for the detection of TP53 mutations in clinical specimens.


METHODS
We studied 33 gynecological samples, 11 apparently normal samples and 22 malignant tumors of various origins. Mutations were identified using single-strand conformational polymorphism analysis with both slab gel and capillary electrophoresis. All DNA samples were amplified with fluorescently labeled PCR primers specific for exons 5-9 for mutation detection.


RESULTS
Of the 33 patient samples tested, mutations and polymorphisms were found in six specimens in three of the five exons scanned; no mutations were found in exons 7 or 9. Both a mutation and polymorphism were found in non-malignant specimens from the control group. The mutations were confirmed by DNA sequence analysis of the regions scanned.


CONCLUSIONS
Mutations and polymorphisms were detected in the clinical samples. All of the mutations were silent except for one non-conservative mutation in exon 5, codon 181. This study demonstrates the usefulness of the National Institute of Standards and Technology (NIST) TP53 reference panel in TP53 mutation detection in clinical tissue specimens.
Sunar-Reeder
Bulbin
B
Department of Medical Biology and Genetics, Faculty of Medicine, Inönü University, Malatya, Turkey.
Atha
Donald H
DH
Aydemir
Songul
S
Reeder
Dennis J
DJ
Tully
Lois
L
Khan
Abraham R
AR
O'Connell
Catherine D
CD
D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000592:standards*; D004273:DNA, Neoplasm / Q000032:analysis; D019075:Electrophoresis, Capillary; D005091:Exons / Q000235:genetics; D005260:Female; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D009369:Neoplasms / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D012015:Reference Standards; D017422:Sequence Analysis, DNA
D004273:DNA, Neoplasm