Non-isotopic silver-stained SSCP is more sensitive than automated direct sequencing for the detection of PTEN mutations in a mixture of DNA extracted from normal and tumor cells

  • Authors:
    • X. Fan
    • F. B. Furnari
    • W. K. Cavenee
    • J. S. Castresana
  • View Affiliations

  • Published online on: May 1, 2001     https://doi.org/10.3892/ijo.18.5.1023
  • Pages: 1023-1026
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Abstract

The sensitivity of non-isotopic PCR-SSCP was compared to direct sequencing of PTEN exons. DNA from leukocytes derived from healthy donors, and from the glioblastoma cell line LN319 was extracted and mixed in different proportions from 0 to 100%. The LN319 cell line contains a point mutation at codon 15 exon 1 of the PTEN gene. The PCR-SSCP experiments demonstrated mutations in samples containing as little as 10% tumor DNA. In contrast, direct DNA sequencing experiments were less sensitive, requiring 30-70% of tumor DNA in the sample, depending on the DNA strand sequenced. In conclusion, PCR-SSCP, in our hands, is more sensitive than automated sequencing for detecting PTEN point mutations. We recommend to always sequence both strands, and take into account that samples containing less than 30% tumor cells should not only be sequenced, but also studied by PCR-SSCP in order to discriminate false negative results.

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May 2001
Volume 18 Issue 5

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Fan X, Furnari FB, Cavenee WK and Castresana JS: Non-isotopic silver-stained SSCP is more sensitive than automated direct sequencing for the detection of PTEN mutations in a mixture of DNA extracted from normal and tumor cells. Int J Oncol 18: 1023-1026, 2001.
APA
Fan, X., Furnari, F.B., Cavenee, W.K., & Castresana, J.S. (2001). Non-isotopic silver-stained SSCP is more sensitive than automated direct sequencing for the detection of PTEN mutations in a mixture of DNA extracted from normal and tumor cells. International Journal of Oncology, 18, 1023-1026. https://doi.org/10.3892/ijo.18.5.1023
MLA
Fan, X., Furnari, F. B., Cavenee, W. K., Castresana, J. S."Non-isotopic silver-stained SSCP is more sensitive than automated direct sequencing for the detection of PTEN mutations in a mixture of DNA extracted from normal and tumor cells". International Journal of Oncology 18.5 (2001): 1023-1026.
Chicago
Fan, X., Furnari, F. B., Cavenee, W. K., Castresana, J. S."Non-isotopic silver-stained SSCP is more sensitive than automated direct sequencing for the detection of PTEN mutations in a mixture of DNA extracted from normal and tumor cells". International Journal of Oncology 18, no. 5 (2001): 1023-1026. https://doi.org/10.3892/ijo.18.5.1023