학술논문
Rapid Detection of SARS-CoV-2 RNA Using Reverse Transcription Recombinase Polymerase Amplification (RT-RPA) with Lateral Flow for N-Protein Gene and Variant-Specific Deletion–Insertion Mutation in S-Protein Gene
Document Type
article
Author
Jose L. Malaga; Monica J. Pajuelo; Michiko Okamoto; Emmanuel Kagning Tsinda; Kanako Otani; Pablo Tsukayama; Lucero Mascaro; Diego Cuicapuza; Masamichi Katsumi; Kazuhisa Kawamura; Hidekazu Nishimura; Akie Sakagami; Yo Ueki; Suguru Omiya; Satoshi Okamoto; Asami Nakayama; Shin-ichi Fujimaki; Chuyao Yu; Sikandar Azam; Eiichi Kodama; Clyde Dapat; Hitoshi Oshitani; Mayuko Saito
Source
Viruses, Vol 15, Iss 6, p 1254 (2023)
Subject
Language
English
ISSN
1999-4915
Abstract
Rapid molecular testing for severe acute respiratory coronavirus 2 (SARS-CoV-2) variants may contribute to the development of public health measures, particularly in resource-limited areas. Reverse transcription recombinase polymerase amplification using a lateral flow assay (RT-RPA-LF) allows rapid RNA detection without thermal cyclers. In this study, we developed two assays to detect SARS-CoV-2 nucleocapsid (N) gene and Omicron BA.1 spike (S) gene-specific deletion–insertion mutations (del211/ins214). Both tests had a detection limit of 10 copies/µL in vitro and the detection time was approximately 35 min from incubation to detection. The sensitivities of SARS-CoV-2 (N) RT-RPA-LF by viral load categories were 100% for clinical samples with high (>9015.7 copies/µL, cycle quantification (Cq): < 25) and moderate (385.5–9015.7 copies/µL, Cq: 25–29.9) viral load, 83.3% for low (16.5–385.5 copies/µL, Cq: 30–34.9), and 14.3% for very low (