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Clinical Chemistry 43: 2251-2255, 1997;
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(Clinical Chemistry. 1997;43:2251-2255.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Single-strand conformation polymorphism analysis to detect p53 mutations: characterization and development of controls

Judith A. Welsh1, Katariina Castrén2 and Kirsi H. Vähäkangas2,a

1 Laboratory of Human Carcinogenesis, NCI, National Institutes of Health, Bethesda, MD 20892 (address for reprint requests: fax 301-496-0497; e-mail welshj{at}intra.nci.nih.gov)

2 Department of Pharmacology and Toxicology, University of Oulu, FIN-90220 Oulu, Finland.
a Author for correspondence. Fax 358-8-537-5247; e-mail kirsi. vahakangas{at}oulu.fi

Single-strand conformation polymorphism (SSCP) analysis is widely used to prescreen mutations in p53 gene. However, standardization of SSCP to detect p53 mutations has rarely been pursued so far. We have developed complete conditions for a temperature-controlled nonradioactive SSCP for mutation detection in amplified p53 exons 4-8, where mutations frequently occur in human tumors. Easily obtainable and clearly distinguishable positive controls were developed by replacing the regular 5' primers in amplification with primers that include one to three mutated sites. Careful purification of the amplified products by gel electrophoresis appeared to be essential. The efficiency of the method was studied by using previously sequenced samples with p53 mutations and the various positive controls. The use of two temperatures (exon 4: 4 °C and 15 °C; other exons: 4 °C and 20 °C) in combination with other optimized conditions resulted in 98% efficiency in mutation detection, which was considered sufficient for routine screening.




The following articles in journals at HighWire Press have cited this article:


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 Hum Exp ToxicolHome page
J Peltonen, J A Welsh, and K H Vahakangas
Is there a role for PCR-SSCP among the methods for missense mutation detection of TP53 gene?
Human and Experimental Toxicology, January 1, 2007; 26(1): 9 - 18.
[Abstract] [PDF]

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 Cancer Res.Home page
N. Hagiwara, L. E. Mechanic, G. E. Trivers, H. L. Cawley, M. Taga, E. D. Bowman, K. Kumamoto, P. He, M. Bernard, S. Doja, et al.
Quantitative Detection of p53 Mutations in Plasma DNA from Tobacco Smokers
Cancer Res., August 15, 2006; 66(16): 8309 - 8317.
[Abstract] [Full Text] [PDF]

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 CarcinogenesisHome page
L. E. Mechanic, A. J. Marrogi, J. A. Welsh, E. D. Bowman, M. A. Khan, L. Enewold, Y.-L. Zheng, S. Chanock, P. G. Shields, and C. C. Harris
Polymorphisms in XPD and TP53 and mutation in human lung cancer
Carcinogenesis, March 1, 2005; 26(3): 597 - 604.
[Abstract] [Full Text] [PDF]

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 J. Virol.Home page
J. S. Hall, R. French, T. J. Morris, and D. C. Stenger
Structure and Temporal Dynamics of Populations within Wheat Streak Mosaic Virus Isolates
J. Virol., November 1, 2001; 75(21): 10231 - 10243.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
K. H. Vahakangas, W. P. Bennett, K. Castren, J. A. Welsh, M. A. Khan, B. Blomeke, M. C. R. Alavanja, and C. C. Harris
p53 and K-ras Mutations in Lung Cancers from Former and Never-Smoking Women
Cancer Res., June 1, 2001; 61(11): 4350 - 4356.
[Abstract] [Full Text] [PDF]

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 GeneticsHome page
J. Jaruzelska, E. Zietkiewicz, M. Batzer, D. E. C. Cole, J.-P. Moisan, R. Scozzari, S. Tavaré, and D. Labuda
Spatial and Temporal Distribution of the Neutral Polymorphisms in the Last ZFX Intron: Analysis of the Haplotype Structure and Genealogy
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Copyright © 1997 by the American Association for Clinical Chemistry.