TY - JOUR
AU - Dong, Yajuan
AU - Zhao, Yongjuan
AU - Li, Shenwei
AU - Wan, Zhenzhou
AU - Lu, Renfei
AU - Yang, Xianguang
AU - Yu, Guoying
AU - Reboud, Julien
AU - Cooper, Jonathan M.
AU - Tian, Zhengan
AU - Zhang, Chiyu
PY - 2022
DA - February
TI - Multiplex, variant-tolerant, real-time RT-LAMP for SARS-CoV-2 detection using HFman probe
JO - ACS Sensors
EP - 739
VL - 7
DO - http://dx.doi.org/10.1021/acssensors.1c02079
AB - Viral evolution impacts diagnostic test performance through the emergence of variants with sequences affecting the efficiency of primer binding. Such variants that evade detection by nucleic acid-based tests are subject to selective pressure, enabling them to spread more efficiently. Here, we report a variant-tolerant diagnostic test for SARS-CoV-2 using a loop-mediated isothermal nucleic acid-based amplification (LAMP) assay containing high-fidelity DNA polymerase and a high-fidelity DNA polymerase-medicated probe (HFman probe). In addition to demonstrating a high tolerance to variable SARS-CoV-2 viral sequences, the mechanism also overcomes frequently observed limitations of LAMP assays arising from non-specific amplification within multiplexed reactions performed in a single “pot”. Results showed excellent clinical performance (sensitivity 94.5%, specificity 100%, n = 190) when compared directly to a commercial gold standard reverse transcription quantitative polymerase chain reaction assay for the extracted RNA from nasopharyngeal samples and the capability of detecting a wide range of sequences containing at least alpha and delta variants. To further validate the test with no sample processing, directly from nasopharyngeal swabs, we also detected SARS-CoV-2 in positive clinical samples (n = 49), opening up the possibility for the assay’s use in decentralized testing.
PB - American Chemical Society
UR - http://eprints.gla.ac.uk/265002/
KW - Coronavirus (COVID-19)
ER