| Info. | Vol.15 - No.4 (2021.12.20) |
|---|---|
| Title | POCT Detection of 14 Respiratory Viruses Using Multiplex RT-PCR |
| Authors | Ji Su Lee1, Jeong Jin Ahn2, Seung Jun Kim2, So Yeon Yu1, Eun Jung Koh3, Seung Hwan Kim3, Heung Sup Sung4, Jin Won Huh5 & Seung Yong Hwang1,* |
| Institutions | 1Department of Molecular and Life Science, Hanyang University, 55, Hanyangdeahak-ro, Sangnok-gu, Ansan, Gyeonggi-do 15588, Republic of Korea 2GeneTrigger, 20, Magokjungang 5-ro 1-gil, Gangseo-gu, Seoul, Republic of Korea 3Department of Bio-Nanotechnology, Hanyang University, 55, Hanyangdeahak-ro, Sangnok-gu, Ansan, Gyeonggi-do 15588, Republic of Korea 4Department of Laboratory Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea 5Department of Pulmonary and Critical Care Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea |
| Abstract | Over the past 6 years, acute respiratory infections have constituted an average of more than 70,000 cases in South Korea. It results in a high mortality rate in infants and the elderly with weak immunity. There are several types of respiratory viruses that invade the human respiratory tract and cause infectious disease. Reverse transcription PCR (RT-PCR) is mainly used for respiratory virus detection owing to its high sensitivity and reproducibility. In response, a multiplex real-time RT-PCR (rRT-PCR) assay was developed for the detection of influenza A and B viruses, parainfluenza viruses 1–4 (PIV1-4), human metapneumovirus, adenovirus, human rhinovirus, respiratory syncytial virus (RSV), and SARS-CoV-2. Detection ability of RT-PCR assay was confirmed by applying it to a portable device capable of point-of-care testing (POCT). Amplicons were synthesized using primer pairs and probe sets designed for each target virus, and a standard curve was constructed to confirm the limit of detection. An experiment using nasopharyngeal swab samples was conducted to understand the field applicability of the rRT-PCR assay. Detection was confirmed in most samples. This study demonstrated that rapid and accurate detection results can be obtained using the multiplex rRT-PCR based POC test, and that it is possible to detect 14 types of respiratory viruses that are generally difficult to distinguish at the same time, enabling timely treatment. Furthermore, we expect that the portable PCR device can significantly reduce the processing procedure of clinical samples before testing, which is the main disadvantage of common RT-PCR tests and can help reduce costs. |
| Keyword | Respiratory virus, RT-PCR, Point-of-care, Molecular diagnosis, SARS-CoV-2 |
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