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Rapid and Sensitive Real-Time Polymerase Chain Reaction Method for Detection and Quantification of 3243A>G Mitochondrial Point Mutation  Rinki Singh,

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Presentation on theme: "Rapid and Sensitive Real-Time Polymerase Chain Reaction Method for Detection and Quantification of 3243A>G Mitochondrial Point Mutation  Rinki Singh,"— Presentation transcript:

1 Rapid and Sensitive Real-Time Polymerase Chain Reaction Method for Detection and Quantification of 3243A>G Mitochondrial Point Mutation  Rinki Singh, Sian Ellard, Andrew Hattersley, Lorna W. Harries  The Journal of Molecular Diagnostics  Volume 8, Issue 2, Pages (May 2006) DOI: /jmoldx Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

2 Figure 1 Crossing points of wild-type and mutant alleles in strong positive (heteroplasmy level, 40%) (a), weak positive (heteroplasmy level, 9%) (b), and normal samples (c). The Journal of Molecular Diagnostics 2006 8, DOI: ( /jmoldx ) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

3 Figure 2 Standard curves generated with plasmids containing 100% mutant and 100% wild-type sequences. The Journal of Molecular Diagnostics 2006 8, DOI: ( /jmoldx ) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

4 Figure 3 Mixing experiment showing limit of detection of the real-time PCR assay. 1, 50% mutant; 2, 40% mutant; 3, 30% mutant; 4, 20% mutant; 5, 10% mutant; 6, 1% mutant; 7, 0.1% mutant; 8, 0.01% mutant. The Journal of Molecular Diagnostics 2006 8, DOI: ( /jmoldx ) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

5 Figure 4 Results obtained with mutant probe tested on triplicate samples of 100% normal plasmids, a normal patient, and a case of known low positive m.3243A>G (9% heteroplasmy). The Journal of Molecular Diagnostics 2006 8, DOI: ( /jmoldx ) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

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