Use ofTP53Reference Materials to Validate Mutations in Clinical Tissue Specimens by Single-Strand Conformational Polymorphism Analysis
作者:
Bulbin Sunar-Reeder,
Donald H Atha,
Songul Aydemir,
Dennis J Reeder,
Lois Tully,
Abraham R Khan,
Catherine D O’Connell,
期刊:
Molecular Diagnosis
(ADIS Available online 2004)
卷期:
Volume 8,
issue 2
页码: 123-130
ISSN:1084-8592
年代: 2004
出版商: ADIS
关键词: Genetic polymorphism;Diagnostic tests
数据来源: ADIS
摘要:
BackgroundAs 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 ofTP53mutations.AimThe goal of this study was to validate the reference materials in providing quality assurance for the detection ofTP53mutations in clinical specimens.MethodsWe 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.ResultsOf 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.ConclusionsMutations 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)TP53reference panel inTP53mutation detection in clinical tissue specimens.
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