Kim, Jung Min Jung, Seungwon Jeon, Eui Ju Kim, Bong Kyun No, Jin Yong Kim, Myung Jong Kim, Heesuk Song, Chang Seon Kim, Sang Kyung 2024-01-19T14:04:17Z 2024-01-19T14:04:17Z 2022-01-10 2021-07 1944-8244 https://pubs.kist.re.kr/handle/201004/116713 As viruses have been threatening global public health, fast diagnosis has been critical to effective disease management and control. Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) is now widely used as the gold standard for detecting viruses. Although a multiplex assay is essential for identifying virus types and subtypes, the poor multiplicity of RT-qPCR makes it laborious and time-consuming. In this paper, we describe the development of a multiplex RT-qPCR platform with hydrogel microparticles acting as independent reactors in a single reaction. To build target-specific particles, target-specific primers and probes are integrated into the particles in the form of noncovalent composites with boron nitride nanotubes (BNNTs) and carbon nanotubes (CNTs). The thermal release characteristics of DNA, primer, and probe from the composites of primer-BNNT and probe-CNT allow primer and probe to be stored in particles during particle production and to be delivered into the reaction. In addition, BNNT did not absorb but preserved the fluorescent signal, while CNT protected the fluorophore of the probe from the free radicals present during particle production. Bicompartmental primer-incorporated network (bcPIN) particles were designed to harness the distinctive properties of two nanomaterials. The bcPIN particles showed a high RT-qPCR efficiency of over 90% and effective suppression of non-specific reactions. 16-plex RT-qPCR has been achieved simply by recruiting differently coded bcPIN particles for each target. As a proof of concept, multiplex one-step RT-qPCR was successfully demonstrated with a simple reaction protocol. English American Chemical Society Highly Selective Multiplex Quantitative Polymerase Chain Reaction with a Nanomaterial Composite Hydrogel for Precise Diagnosis of Viral Infection Article 10.1021/acsami.1c03434 1 ACS Applied Materials & Interfaces, v.13, no.26, pp.30295 - 30305 ACS Applied Materials & Interfaces 13 26 30295 30305 scie scopus 000672492800005 2-s2.0-85110685174 Nanoscience & Nanotechnology Materials Science, Multidisciplinary Science & Technology - Other Topics Materials Science Article BORON-NITRIDE NANOTUBES RT-PCR DNA GRAPHENE DESORPTION PRODUCT DESIGN ARRAY virus detection reverse-transcription quantitative PCR boron nitride nanotube (BNNT) carbon nanotube (CNT) hydrogel multiplex assay