ABSTRACT LamPORE is a novel diagnostic platform for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA combining loop-mediated isothermal amplification with nanopore sequencing, which could potentially be used to analyze thousands of samples per day on a single instrument. We evaluated the performance of LamPORE against reverse transcriptase PCR (RT-PCR) using RNA extracted from spiked respiratory samples and stored nose and throat swabs collected at two UK hospitals. The limit of detection of LamPORE was 10 genome copies/μl of extracted RNA, which is above the limit achievable by RT-PCR, but was not associated with a significant reduction of sensitivity in clinical samples. Positive clinical specimens came mostly from patients with acute symptomatic infection, and among them, LamPORE had a diagnostic sensitivity of 99.1% (226/228; 95% confidence interval [CI], 96.9% to 99.9%). Among negative clinical specimens, including 153 with other respiratory pathogens detected, LamPORE had a diagnostic specificity of 99.6% (278/279; 98.0% to 100.0%). Overall, 1.4% (7/514; 0.5% to 2.9%) of samples produced an indeterminate result on first testing, and repeat LamPORE testing on the same RNA extract had a reproducibility of 96.8% (478/494; 94.8% to 98.1%). LamPORE has a similar performance as RT-PCR for the diagnosis of SARS-CoV-2 infection in symptomatic patients and offers a promising approach to high-throughput testing.
【저자키워드】 SARS-CoV-2, Diagnosis, nanopore sequencing, LamPORE, 【초록키워드】 coronavirus, SARS-COV-2 infection, Nanopore, Sequencing, Genome, Infection, diagnostic, severe acute respiratory syndrome Coronavirus, RT-PCR, RNA, clinical samples, sensitivity, specificity, Symptomatic patients, Loop-mediated isothermal amplification, diagnostic sensitivity, Patient, limit of detection, nanopore sequencing, Pathogens, isothermal amplification, Swab, respiratory, reverse transcriptase PCR, specimens, platform, symptomatic infection, nose and throat, reverse transcriptase, diagnostic specificity, reduction, acute respiratory syndrome, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, 95% confidence interval, throat swabs, respiratory pathogen, specimen, reproducibility, extracted RNA, offer, UK hospitals, approach, limit, respiratory sample, produced, collected, evaluated, per day, diagnosis of SARS-CoV-2, symptomatic patient, 【제목키워드】 Loop-mediated isothermal amplification,