ArticlePloS one2016
Novel Methodology for Rapid Detection of KRAS Mutation Using PNA-LNA Mediated Loop-Mediated Isothermal Amplification.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 81 citations in OpenAlex.
- Cancer theranostics.Nature reviews. Cancer · 2026Review
- Bst DNA Polymerase: Structure, Properties and Engineering Strategies in LAMP.International journal of molecular sciences · 2026Review
- Advancements in somatic mutation detection through loop-mediated isothermal amplification (LAMP) in human tumors.Journal of applied genetics · 2026Review
- A point-of-care assay for genotyping women prior to ovarian stimulation with recombinant or human menopausal gonadotropins in assisted reproduction.Frontiers in endocrinology · 2026Article
- Review
- Integration of Point-of-Care Technology in the Decoding Process of Single Nucleotide Polymorphism for Healthcare ApplicationMicromachines · 2025Review
- Advances in nucleic acid probe-based detection of gene point mutations: a review.Frontiers in chemistry · 2025Review
- KRAS mutations detection methodology: from RFLP to CRISPR/Cas based methods.Functional & integrative genomics · 2024Review
- Versatility of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) from diagnosis of early pathological infection to mutation detection in organisms.Molecular biology reports · 2024Review
- RapidBiology methods & protocols · 2024Article
- Development of a Detection System forDiagnostics (Basel, Switzerland) · 2023Article
- Vivid COVID-19 LAMP is an ultrasensitive, quadruplexed test using LNA-modified primers and a zinc ion and 5-Br-PAPS colorimetric detection system.Communications biology · 2023Article
- Article
- Programmable design of isothermal nucleic acid diagnostic assays through abstraction-based models.Nature communications · 2022Article
- Cationic copolymers that enhance wild-type-specific suppression in BNA-clamp PCR and preferentially increase theBiology methods & protocols · 2022Article
- Loop-Mediated Isothermal Amplification: From Theory to Practice.Russian journal of bioorganic chemistry · 2022Article
- Design of Oligonucleotides for Allele-Specific Amplification Based on PCR and Isothermal Techniques.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Allele-Specific Recombinase Polymerase Amplification to Detect Sickle Cell Disease in Low-Resource Settings.Analytical chemistry · 2021Article
- Review
- Marker-Assisted Selection for Pollen-Free Somatic Plants of Sugi (Japanese Cedar,Frontiers in plant science · 2021Article
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Detecting point mutation of human cancer cells quickly and accurately is gaining in importance for pathological diagnosis and choice of therapeutic approach. In the present study, we present novel methodology, peptide nucleic acid-locked nucleic acid mediated loop-mediated isothermal amplification (PNA-LNA mediated LAMP), for rapid detection of KRAS mutation using advantages of both artificial DNA and LAMP. PNA-LNA mediated LAMP reactions occurred under isothermal temperature conditions of with 4 primary primers set for the target regions on the KRAS gene, clamping PNA probe that was complimentary to the wild type sequence and LNA primers complementary to the mutated sequences. PNA-LNA mediated LAMP was applied for cDNA from 4 kinds of pancreatic carcinoma cell lines with or without KRAS point mutation. The amplified DNA products were verified by naked-eye as well as a real-time PCR equipment. By PNA-LNA mediated LAMP, amplification of wild type KRAS DNA was blocked by clamping PNA probe, whereas, mutant type KRAS DNA was significantly amplified within 50 min. Mutant alleles could be detected in samples which diluted until 0.1% of mutant-to-wild type ratio. On the other hand, mutant alleles could be reproducibly with a mutant-to-wild type ratio of 30% by direct sequencing and of 1% by PNA-clamping PCR. The limit of detection (LOD) of PNA-LNA mediated LAMP was much lower than the other conventional methods. Competition of LNA clamping primers complementary to two different subtypes (G12D and G12V) of mutant KRAS gene indicated different amplification time depend on subtypes of mutant cDNA. PNA-LNA mediated LAMP is a simple, rapid, specific and sensitive methodology for the detection of KRAS mutation.
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