B-232 Versatile, Extraction-Free Nucleic Acid Sample Preparation in 15 Seconds Via Hyperbaric Heating

• DOI: 10.1093/clinchem/hvad097.558
• Publication Date: 2023-09-27T23:24:20Z
• Authors: J A Hambalek; R L Martin; E Alvarado; P W Halle; R E Davis; H Janwari; H Kido; R Peytavi
• Keywords: Sample Preparation

Abstract Nucleic acid extraction-based PCR sample prep is currently limited by long processing times, liquid transfers, kit cost, and lab storage requirements. Extraction-free approaches enable streamlined prep, but lack sensitivity compared to methods. We studied several types demonstrate the effectiveness of a novel 15-second Hyperbaric Heating (HBH) process prepare nucleic acids for PCR. HBH exposes short duration, high temperature environment effectively lyse organisms denature inhibitors such as nucleases in seconds. Samples are introduced sealed pressure-tight metallic vessel, which then heated seconds using magnetic induction. Within vaporization aqueous raises internal pressure, enabling an higher than 100°C be achieved. Chelating agents other proteins also protect target from fragmentation. Post HBH, ready direct amplification without further dilution or extraction. Efficacy efficient was tested against commercial kits. applied viral, bacterial, eukaryotic, fungal organisms. For SARS-CoV-2, detected down 250 viral copies/mL. Additionally, outperformed kits Candida albicans Bacillus subtilis. generalizable technology with comparable that current extraction have multitude demonstrated can employed rapid comprehensive approach applications involve post prep. Future integration devices suitable point care molecular testing underway. envision useful, method where simplicity, speed essential preparing amplification.