Kifaro, Emmanuel George Kim Mi Jung Jung, Seungwon Jang, Yoon-ha Moon, Sungyeon Lee, Dong-Hun Song, Chang-Seon Misinzo, Gerald Kim, Sang Kyung 2024-01-19T10:30:45Z 2024-01-19T10:30:45Z 2023-02-10 2023-01 https://pubs.kist.re.kr/handle/201004/114132 Since its discovery, polymerase chain reaction (PCR) has emerged as an important technology for the diagnosis and identification of infectious diseases. It is a highly sensitive and reliable nucleic acids (NA) detection tool for various sample types. However, stool, which carries the most abundant micro-organisms and physiological byproducts, remains to be the trickiest clinical specimen for molecular detection of pathogens. Herein, we demonstrate the novel application of hydrogel microparticles as carriers of viral RNA from stool samples without prior RNA purification for real-time polymerase chain reaction (qPCR). In each microparticle of primer-incorporated network (PIN) as a self-sufficient reaction compartment, immobilized reverse transcription (RT) primers capture the viral RNA by hybridization and directly initiate RT of RNA to generate a pool of complementary DNA (PIN-cDNA pool). Through a simple operation with a portable thermostat device, a PIN-cDNA pool for influenza A virus (IAV) was obtained in 20 min. The PIN-cDNA pools can be stored at room temperature, or directly used to deliver cDNA templates for qPCR. The viral cDNA templates were freely released in the subsequent qPCR to allow amplification efficiency of over 91%. The assay displayed good linearity, repeatability, and comparable limit of detection (LoD) with a commercialized viral RNA purification kit. As a proof of concept, this technology carries a huge potential for onsite application to improve human and animal infectious disease surveillance activities using stool samples without the need for a laboratory or centrifuge for sample preparation. English MDPI AG Microparticles as Viral RNA Carriers from Stool for Stable and Sensitive Surveillance Article 10.3390/diagnostics13020261 1 Diagnostics, v.13, no.2 Diagnostics 13 2 Y scie scopus 000917311200001 2-s2.0-85146789431 Medicine, General & Internal General & Internal Medicine Article AVIAN INFLUENZA-VIRUS DNA EXTRACTION FECAL SAMPLES PCR BACTERIAL COMBINATION COLLECTION INHIBITORS DIAGNOSIS hydrogel microparticles viral RNA carrier qPCR influenza A virus stool samples