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Innovative PNA-LB mediated allele-specific LAMP for KRAS mutation profiling on a compact lab-on-a-disc device.
- Source :
-
Talanta . Aug2024, Vol. 276, pN.PAG-N.PAG. 1p. - Publication Year :
- 2024
-
Abstract
- Tailored healthcare, an approach focused on individual patients, requires integrating emerging interdisciplinary technologies to develop accurate and user-friendly diagnostic tools. KRAS mutations, prevalent in various common cancers, are crucial determinants in selecting patients for novel KRAS inhibitor therapies. This study presents a novel state-of-the-art Lab-on-a-Disc system utilizing peptide nucleic acids-loop backward (PNA-LB) mediated allele-specific loop-mediated isothermal amplification (LAMP) for detecting the frequent G12D KRAS mutation, signifying its superiority over alternative mutation detection approaches. The designed Lab-on-a-Disc system demonstrated exceptional preclinical and technical precision, accuracy, and versatility. By applying varying cutoff values to PNA- LB LAMP reactions, the assay's sensitivity and specificity were increased by 80 % and 90 %, respectively. The device's key advantages include a robust microfluidic Lab-on-a-Disc design, precise rotary control, and a cutting-edge induction heating module. These features enable multiplexing of LAMP reactions with high reproducibility and repeatability, with CV% values less than 3.5 % and 5.5 %, respectively. The device offers several methods for accurate endpoint result detection, including naked-eye observation, RGB image analysis using Python code, and time of fluorescence (T f) values. Preclinical specificity and sensitivity, assessed using different cutoffs for Eva-Green fluorescence T f values and pH-sensitive dyes, demonstrated comparable performance to the best standard methods. Overall, this study represents a significant step towards tailoring treatment strategies for cancer patients through precise and efficient mutation detection technologies. [Display omitted] • New Lab-on-a-Disc system uses PNA/LNA in LAMP for precise, accurate detection of frequent G12D KRAS mutation. • Increased sensitivity (80%) and specificity (90%) via varied cut-off values for LNA-PNA LAMP reactions. • Multiple endpoint result detection methods: naked-eye observation, RGB image analysis, and fluorescence Tf values. • Crafting a reliable microfluidic disc with rotary control and advanced induction heating ensures high reproducibility (CV% < 3.5% and 5.5%). • Suitable for point-of-care diagnostics and compact lab devices, detecting multiple mutations, personalized medicine. [ABSTRACT FROM AUTHOR]
- Subjects :
- *RAS oncogenes
*INDUCTION heating
*LAMPS
*PEPTIDE nucleic acids
*IMAGE analysis
Subjects
Details
- Language :
- English
- ISSN :
- 00399140
- Volume :
- 276
- Database :
- Academic Search Index
- Journal :
- Talanta
- Publication Type :
- Academic Journal
- Accession number :
- 177747382
- Full Text :
- https://doi.org/10.1016/j.talanta.2024.126224