The IDH1 mutation is an important diagnostic and prognostic biomarker used to characterize glioblastoma (GBM). Patients harboring the IDH1 mutation have improved overall survival following maximal resection. Knowledge of the IDH1 mutation status allows the surgeon to modify the surgical plan; however, no existing molecular test can provide this information intraoperatively. We designed a novel colorimetric peptide nucleic acid loop-mediated isothermal amplification (PNA-LAMP) method that rapidly detects the IDH1 R132H mutation in GBM. PNA-LAMP amplifies target DNA under isothermal conditions with high specificity and speed. The PNA prevents amplification of wild-type IDH1 DNA, while allowing amplification of the R132H variant if present. We used a pH-sensitive colorimetric detection method for visual determination of amplification in under one hour. Characterization of the assay was performed with plasmid DNA containing the IDH1 wild-type and R132H variant sequences. Amplification was confirmed using gel electrophoresis, and this analysis suggests that the assay is more sensitive than Sanger sequencing – the gold standard for IDH1 mutation identification. This study is the first to attempt to develop a colorimetric LAMP assay for GBM tumor characterization, and only the third application of the PNA-LAMP method to detect acquired mutations in cancer. This novel molecular assay is a simple, specific, and rapid way to identify the presence of the IDH1 R132H variant associated with GBM.
","abstract_html":"<p>The <em>IDH1</em> mutation is an important diagnostic and prognostic biomarker used to characterize glioblastoma (GBM). Patients harboring the <em>IDH1</em> mutation have improved overall survival following maximal resection. Knowledge of the <em>IDH1</em> mutation status allows the surgeon to modify the surgical plan; however, no existing molecular test can provide this information intraoperatively. We designed a novel colorimetric peptide nucleic acid loop-mediated isothermal amplification (PNA-LAMP) method that rapidly detects the <em>IDH1</em> R132H mutation in GBM. PNA-LAMP amplifies target DNA under isothermal conditions with high specificity and speed. The PNA prevents amplification of wild-type <em>IDH1</em> DNA, while allowing amplification of the R132H variant if present. We used a pH-sensitive colorimetric detection method for visual determination of amplification in under one hour. Characterization of the assay was performed with plasmid DNA containing the <em>IDH1</em> wild-type and R132H variant sequences. Amplification was confirmed using gel electrophoresis, and this analysis suggests that the assay is more sensitive than Sanger sequencing – the gold standard for <em>IDH1</em> mutation identification. This study is the first to attempt to develop a colorimetric LAMP assay for GBM tumor characterization, and only the third application of the PNA-LAMP method to detect acquired mutations in cancer. This novel molecular assay is a simple, specific, and rapid way to identify the presence of the <em>IDH1</em> R132H variant associated with GBM. </p>","abstract_has_math":false,"creators":["Raack, Edward J"],"institution":null,"degree_name":"Master of Science","degree_level":"Thesis","degree_discipline":"Clinical Sciences","degree_department":null,"school":null,"contributors":["Dr. Paul Mann"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2017,"date_issued":"2017-07-01T07:00:00Z","date_published":"2017-07-01T07:00:00Z","updated_at":"2026-07-24T03:23:47Z","subjects":["Loop-mediated isothermal amplification","glioblastoma","isocitrate dehydrogenase","molecular genetics","assay development","Biotechnology","Genetics","Genetics and Genomics"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://commons.nmu.edu/theses/158","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Dr. Paul Mann"]},{"key":"dc:creator","label":"Author","values":["Raack, Edward J"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2017-07-10T07:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Clinical Sciences"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Loop-mediated isothermal amplification","glioblastoma","isocitrate dehydrogenase","molecular genetics","assay development","Biotechnology","Genetics","Genetics and Genomics"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://commons.nmu.edu/theses/158"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The IDH1 mutation is an important diagnostic and prognostic biomarker used to characterize glioblastoma (GBM). Patients harboring the IDH1 mutation have improved overall survival following maximal resection. Knowledge of the IDH1 mutation status allows the surgeon to modify the surgical plan; however, no existing molecular test can provide this information intraoperatively. We designed a novel colorimetric peptide nucleic acid loop-mediated isothermal amplification (PNA-LAMP) method that rapidly detects the IDH1 R132H mutation in GBM. PNA-LAMP amplifies target DNA under isothermal conditions with high specificity and speed. The PNA prevents amplification of wild-type IDH1 DNA, while allowing amplification of the R132H variant if present. We used a pH-sensitive colorimetric detection method for visual determination of amplification in under one hour. Characterization of the assay was performed with plasmid DNA containing the IDH1 wild-type and R132H variant sequences. Amplification was confirmed using gel electrophoresis, and this analysis suggests that the assay is more sensitive than Sanger sequencing – the gold standard for IDH1 mutation identification. This study is the first to attempt to develop a colorimetric LAMP assay for GBM tumor characterization, and only the third application of the PNA-LAMP method to detect acquired mutations in cancer. This novel molecular assay is a simple, specific, and rapid way to identify the presence of the IDH1 R132H variant associated with GBM.
"]},{"key":"dc:title","label":"Title","values":["A Rapid Colorimetric Peptide Nucleic Acid Loop-Mediated Isothermal Amplification Assay for the Detection of the IDH1 Mutation in Glioblastoma"]}]}],"canonical_facts":{"dc:contributor":["Dr. Paul Mann"],"dc:creator":["Raack, Edward J"],"dc:date.available":["2017-07-10T07:00:00Z"],"dc:description.abstract":["The IDH1 mutation is an important diagnostic and prognostic biomarker used to characterize glioblastoma (GBM). Patients harboring the IDH1 mutation have improved overall survival following maximal resection. Knowledge of the IDH1 mutation status allows the surgeon to modify the surgical plan; however, no existing molecular test can provide this information intraoperatively. We designed a novel colorimetric peptide nucleic acid loop-mediated isothermal amplification (PNA-LAMP) method that rapidly detects the IDH1 R132H mutation in GBM. PNA-LAMP amplifies target DNA under isothermal conditions with high specificity and speed. The PNA prevents amplification of wild-type IDH1 DNA, while allowing amplification of the R132H variant if present. We used a pH-sensitive colorimetric detection method for visual determination of amplification in under one hour. Characterization of the assay was performed with plasmid DNA containing the IDH1 wild-type and R132H variant sequences. Amplification was confirmed using gel electrophoresis, and this analysis suggests that the assay is more sensitive than Sanger sequencing – the gold standard for IDH1 mutation identification. This study is the first to attempt to develop a colorimetric LAMP assay for GBM tumor characterization, and only the third application of the PNA-LAMP method to detect acquired mutations in cancer. This novel molecular assay is a simple, specific, and rapid way to identify the presence of the IDH1 R132H variant associated with GBM.
"],"dc:identifier":["https://commons.nmu.edu/theses/158"],"dc:subject":["Loop-mediated isothermal amplification","glioblastoma","isocitrate dehydrogenase","molecular genetics","assay development","Biotechnology","Genetics","Genetics and Genomics"],"dc:title":["A Rapid Colorimetric Peptide Nucleic Acid Loop-Mediated Isothermal Amplification Assay for the Detection of the IDH1 Mutation in Glioblastoma"],"thesis:degree_discipline":["Clinical Sciences"],"thesis:degree_level":["Thesis"],"thesis:degree_name":["Master of Science"]},"updated_at":"2026-07-24T03:23:47Z"}