Stabilized Exfoliated Black Phosphorus Nanosheets for Fungal DNA-Extraction Assay

Abstract

DNA extraction is essential in fields like medicine, biotechnology, and environmental science. Traditional DNA-extraction methods often face challenges, especially with organisms like Aspergillus that have tough cell walls. We propose a novel method using black phosphorus nanosheets (BPNSs) in NMP(1-Methyl-2-pyrrolidinone)-NaOH solvent with dimethyl suberimidate (DMS) for efficient fungal DNA extraction. BP, with its unique layered structure and photothermal effects, combined with laser irradiation, eliminates thermal steps and hazardous chemicals, simplifying the process. Hence, BPNSs in NMP-NaOH solvent with DMS (BNNDs) are synthesized via a liquid-phase ultrasonic exfoliation method in NMP-NaOH solvent to produce monolayer BP, with the NMP-NaOH serving as the lysis buffer and DMS as the cross-linking agent. This approach significantly improves fungal spore lysis and DNA capture efficiency, achieving fungal DNA isolation in under 15 min with high sensitivity and specificity. Real-time qPCR analysis shows lower threshold cycle values and higher DNA yields compared to commercial kits. This technique is efficient, less labor-intensive, and avoids hazardous chemicals, unlike commercial kits. The assay has high sensitivity, detecting fungal DNA concentrations as low as 101 spores per mL. This BNND method offers a rapid, simple, and highly efficient solution for fungal DNA extraction, promising significant advancements in research and diagnostics across various fields.