바이오 실리카를 이용한 병원체 핵산 분리 기술 연구

Abstract

Diseases caused by pathogens are an important problem worldwide, and infectious diseases caused by zoonosis are increasing the health risks of people in low-income countries. Moreover, diseases diagnosis usually requires complicated pretreatment processes and is hard to obtain Nucleic acid (NA) due to low concentrations and expensive equipment. In this study, we developed a more efficient system for the enrichment of pathogens and isolation of NA by amine-functionalized the surface of the biocompatible nanomaterial diatomaceous earth (DE) and combining it with other molecules to make new compounds or adding chemicals with amine-functionalized diatomaceous earth (ADE). First, a pumpkin-shaped molecular cucurbituril (CB) coating ADE composite (ADE-CB) was fabricated to support effective sample enrichment and NA isolation from enriched pathogens and cells. Using ADE-CB, enrichment of pathogen samples was confirmed to have a concentration efficiency of 90% or more in a short time by simply shaking the hand without any equipment. Next, a syringe filter system was used together with ADE to conveniently and easily enriched pathogens to develop a technology that isolates nucleic acids without equipment in a short period of time. In addition, we developed a technology that can extract multiple samples at once by combining with 96-well filter/membrane plates. Our technology is 10 to 100 times more sensitive than commercial kits and inexpensive due to no need additional experimental equipment. This effective method for capturing pathogen and nucleic acid might be useful for preparing samples for diagnostic systems and several other benefits include the simplicity of producing its components and its ease of operation, and it can be readily integrated with other assays for point-of-care test (POCT).