Cold single-strand conformation polymorphism analysis: Optimization for detection of APC gene mutations in patients with familial adenomatous polyposis

  • Authors:
    • Pierluigi Strippoli
    • Silvia Sarchielli
    • Renato Santucci
    • Gian Paolo Bagnara
    • Giovanni Brandi
    • Guido Biasco
  • View Affiliations

  • Published online on: November 1, 2001     https://doi.org/10.3892/ijmm.8.5.567
  • Pages: 567-572
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Abstract

Over 200 adenomatous polyposis coli (APC) gene mutations have been described in familial adenomatous polyposis (FAP) patients. Recent single-strand conformation polymorphism (SSCP) screening methods have introduced minigel runs, simple ethidium bromide staining and external temperature control without any loss of sensitivity (cold-SSCP). In order to test the effectiveness in APC mutation detection, cold-SSCP was employed following polymerase chain reaction (PCR) amplification in three patients with FAP. Different running parameter combinations were compared. The three mutations already known were all diagnosed by cold-SSCP. The gel concentration was found to be essential in detecting the single-base substitution in fragments of different lengths. The observation of deletions was not affected by gel concentrations and heteroduplex bands were always produced. The temperature or glycerol addition did not significantly affect sensitivity. This modified cold-SSCP method provides a simple and effective way for detecting several known Apc gene mutations without any loss of sensitivity and could be useful for large-scale molecular diagnosis of FAP.

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November 2001
Volume 8 Issue 5

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Strippoli P, Sarchielli S, Santucci R, Bagnara GP, Brandi G and Biasco G: Cold single-strand conformation polymorphism analysis: Optimization for detection of APC gene mutations in patients with familial adenomatous polyposis. Int J Mol Med 8: 567-572, 2001
APA
Strippoli, P., Sarchielli, S., Santucci, R., Bagnara, G.P., Brandi, G., & Biasco, G. (2001). Cold single-strand conformation polymorphism analysis: Optimization for detection of APC gene mutations in patients with familial adenomatous polyposis. International Journal of Molecular Medicine, 8, 567-572. https://doi.org/10.3892/ijmm.8.5.567
MLA
Strippoli, P., Sarchielli, S., Santucci, R., Bagnara, G. P., Brandi, G., Biasco, G."Cold single-strand conformation polymorphism analysis: Optimization for detection of APC gene mutations in patients with familial adenomatous polyposis". International Journal of Molecular Medicine 8.5 (2001): 567-572.
Chicago
Strippoli, P., Sarchielli, S., Santucci, R., Bagnara, G. P., Brandi, G., Biasco, G."Cold single-strand conformation polymorphism analysis: Optimization for detection of APC gene mutations in patients with familial adenomatous polyposis". International Journal of Molecular Medicine 8, no. 5 (2001): 567-572. https://doi.org/10.3892/ijmm.8.5.567