Detection and Discrimination between Deletional and Non-Deletional Prader-Willi and Angelman Syndromes by Methylation-Specific PCR and Quantitative Melting.

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Presentation transcript:

Detection and Discrimination between Deletional and Non-Deletional Prader-Willi and Angelman Syndromes by Methylation-Specific PCR and Quantitative Melting Curve Analysis Wen Wang, Hai-Yang Law, Samuel S. Chong The Journal of Molecular Diagnostics Volume 11, Issue 5, Pages 446-449 (September 2009) DOI: 10.2353/jmoldx.2009.090015 Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 MS-PCR and quantitative MCA analysis of the LIS1 and SNRPN loci. The highlighted columns indicate the LIS1, paternal SNRPN, and maternal SNRPN peaks. Three peaks are observed in a normal sample (A), namely LIS1 (Tm = ∼76.0°C), paternal SNRPN (Tm = ∼83.3°C), and maternal SNRPN (Tm = ∼87.7°C). Only the LIS1 and maternal SNRPN peaks are observed in PWS affected patients (B, C). The maternal SNRPN peak is lower than the LIS1 peak in deletional PWS (B), but higher than LIS1 in non-deletional PWS (C). Only the LIS1 and paternal SNRPN peaks are observed in AS affected patients (D, E). The paternal SNRPN peak is higher than the LIS1 peak in both deletional (D) and non-deletional (E) AS, but the peak height ratio differs between the two mutation groups without overlap. In a sample with paternal SNRPN duplication (F), the LIS1, paternal SNRPN, and maternal SNRPN peaks are observed. However, the paternal SNRPN peak is much higher than the LIS1 peak, as compared with a normal sample. In both (A) and (F), the arrow indicates an additional minor peak lying between the LIS1 and paternal SNRPN peaks, which may have been derived from melting of a heteroduplex of paternal and maternal SNRPN products. The Journal of Molecular Diagnostics 2009 11, 446-449DOI: (10.2353/jmoldx.2009.090015) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Peak height ratios for deletional and non-deletional PWS/AS samples from replicate PCR experiments. Variations in the peak height ratios were observed between runs. However, the ratios in the deletion and non-deletion groups within each run did not overlap. The Journal of Molecular Diagnostics 2009 11, 446-449DOI: (10.2353/jmoldx.2009.090015) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions