Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Novel Genetic Risk Factor Identified for L-Asparaginase-Induced Pancreatitis in Pediatric Patients With Cancer.

Pediatric blood & cancer·2026
Same author

Four Pharmacogenomic Variants Strongly Linked to Corticosteroid-Induced Avascular Necrosis in Children with Cancer.

Journal of clinical pharmacology·2025
Same author

Selecting the Right Technique for the Treatment of Submental Adiposity.

Facial plastic surgery : FPS·2025
Same author

Amino Acid Stress Response Genes Contribute to a 25-Fold Increased Risk of L-Asparaginase-Induced Hypersensitivity.

Pediatric blood & cancer·2025
Same author

A genetically encoded fluorescent sensor enables sensitive and specific detection of IDH mutant associated oncometabolite D-2-hydroxyglutarate.

BMC cancer·2025
Same author

IDH Mutations in Glioma: Molecular, Cellular, Diagnostic, and Clinical Implications.

Biology·2024

Related Experiment Video

Updated: Jun 29, 2025

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
06:18

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1

Published on: March 13, 2018

14.3K

Rapid IDH1-R132 genotyping panel utilizing locked nucleic acid loop-mediated isothermal amplification.

Kristian A Choate1,2, Edward J Raack2,3, Paul B Mann2,3

  • 1Department of Biology, Northern Michigan University, Marquette, MI, United States.

Biology Methods & Protocols
|April 3, 2024
PubMed
Summary
This summary is machine-generated.

This study introduces a rapid genotyping method using locked nucleic acids (LNAs) and loop-mediated isothermal amplification (LAMP) for detecting IDH1-R132 variants. The LNA-LAMP assay provides quick, accurate single-nucleotide variant detection without DNA extraction.

Keywords:
IDH (Isocitrate dehydrogenase)IDH1 R132HLocked Nucleic Acids (LNA)Loop Mediated Isothermal Amplification (LAMP)SNV (single nucleotide variants)SNV discriminationgenotyping panelgliomamutant IDH1 glioma

More Related Videos

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications
07:40

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications

Published on: June 29, 2020

13.4K
Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification
07:25

Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification

Published on: June 25, 2020

6.5K

Related Experiment Videos

Last Updated: Jun 29, 2025

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
06:18

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1

Published on: March 13, 2018

14.3K
Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications
07:40

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications

Published on: June 29, 2020

13.4K
Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification
07:25

Detection of Phytophthora capsici in Irrigation Water using Loop-Mediated Isothermal Amplification

Published on: June 25, 2020

6.5K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Single-nucleotide variants (SNVs) are crucial for understanding human health and disease.
  • Current genotyping methods often involve lengthy procedures, including nucleic acid extraction, and extended analytical times.
  • Rapid and efficient detection of specific SNVs is needed for timely diagnosis and research.

Purpose of the Study:

  • To develop a fast and efficient protocol for allelic discrimination of isocitrate dehydrogenase 1 R132 (IDH1-R132) variants.
  • To integrate locked nucleic acids (LNAs) with loop-mediated isothermal amplification (LAMP) for enhanced genotyping.
  • To evaluate the assay's performance in purified DNA and complex biological samples like tumor lysates.

Main Methods:

  • Utilized self-annealing loop primers incorporating locked nucleic acids (LNAs).
  • Employed loop-mediated isothermal amplification (LAMP) for isothermal DNA amplification and detection.
  • Assessed specificity using synthetic DNA and glioma tumor lysates with known IDH1-R132 mutational status.
  • Evaluated a pH-based colorimetric indicator and an absorbance ratio end-point format for visual and objective interpretation.

Main Results:

  • Demonstrated specific single-nucleotide variant (SNV) discrimination for five IDH1-R132 variants.
  • Achieved genotyping results in approximately 35 minutes directly from glioma tumor lysates, bypassing nucleic acid extraction.
  • Showcased the assay's ability to detect variants in the presence of excess wild-type DNA.
  • Confirmed high reproducibility with no false-positive or false-negative results.

Conclusions:

  • The LNA-LAMP assay offers a rapid, specific, and reproducible method for genotyping IDH1-R132 variants.
  • This protocol eliminates the need for nucleic acid extraction, significantly reducing assay time.
  • The assay is suitable for detecting single base differences in various sample types, including challenging samples with mixed genotypes.