Large Instrument- and Detergent-Free Assay for Ultrasensitive Nucleic Acids Isolation via Binary Nanomaterial

• Published in: Analytical chemistry (Washington) Vol. 90; no. 8; pp. 5108 - 5115
• Main Authors: Liu, Huifang, Zhao, Fei, Jin, Choong Eun, Koo, Bonhan, Lee, Eun Yeong, Zhong, Linlin, Yun, Kyusik, Shin, Yong
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 17.04.2018
• Subjects: Analytical chemistry; Bacteria - genetics; Biological properties; Chemistry; Deoxyribonucleic acid; Detergents; Detergents - chemistry; Diagnostic systems; Diatomaceous earth; Dimethyl Suberimidate - chemistry; DNA; DNA, Bacterial - analysis; DNA, Bacterial - isolation & purification; Endopeptidase K - metabolism; HCT116 Cells; Humans; Medical diagnosis; Molecules; Nanomaterials; Nanostructures - chemistry; Nucleic acids; Nucleic Acids - analysis; Nucleic Acids - isolation & purification; Organic chemistry; Particle Size; Physical properties; Point of care testing; Polymerase Chain Reaction - methods; Proteinase; Ribonucleic acid; RNA; Temperature; Temperature control; Zinc oxide; Zinc Oxide - chemistry
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.7b05136

Nucleic acid-based diagnostics are widely used for clinical applications due to their powerful recognition of biomolecule properties. Isolation and purification of nucleic acids such as DNA and RNA in the diagnostic system have been severely hampered in point-of-care testing because of low recovery yields, degradation of nucleic acids due to the use of chaotropic detergent and high temperature, and the requirement of large instruments such as centrifuges and thermal controllers. Here, we report a novel large instrument- and detergent-free assay via binary nanomaterial for ultrasensitive nucleic acid isolation and detection from cells (eukaryotic and prokaryotic). This binary nanomaterial couples a zinc oxide nanomultigonal shuttle (ZnO NMS) for cell membrane rupture without detergent and temperature control and diatomaceous earth with dimethyl suberimidate complex (DDS) for the capture and isolation of nucleic acids (NA) from cells. The ZnO NMS was synthesized to a size of 500 nm to permit efficient cell lysis at room temperature within 2 min using the biological, chemical, and physical properties of the nanomaterial. By combining the ZnO NMS with the DDS and proteinase K, the nucleic acid extraction could be completed in 15 min with high quantity and quality. For bacterial cells, DNA isolation with the binary nanomaterial yielded 100 times more DNA, than a commercial spin column based reference kit, as determined by the NanoDrop spectrophotometer. We believe that this binary nanomaterial will be a useful tool for rapid and sensitive nucleic acid isolation and detection without large instruments and detergent in the field of molecular diagnostics.