A handheld point-of-care system for rapid detection of SARS-CoV-2 in under 20 minutes

Abstract

The COVID-19 pandemic is a global health emergency characterized by the high rate of transmission and ongoing increase of cases globally. Rapid point-of-care (PoC) diagnostics to detect the causative virus, SARS-CoV-2, are urgently needed to identify and isolate patients, contain its spread and guide clinical management. In this work, we report the development of a rapid PoC diagnostic test (< 20 min) based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) and semiconductor technology for the detection of SARS-CoV-2 from extracted RNA samples. The developed LAMP assay was tested on a real-time benchtop instrument (RT-qLAMP) showing a lower limit of detection of 10 RNA copies per reaction. It was validated against 183 clinical samples including 127 positive samples (screened by the CDC RT-qPCR assay). Results showed 90.55% sensitivity and 100% specificity when compared to RT-qPCR and average positive detection times of 15.45 ± 4.43 min. For validating the incorporation of the RT-LAMP assay onto our PoC platform (RT-eLAMP), a subset of samples was tested (n=40), showing average detection times of 12.89 ± 2.59 min for positive samples (n=34), demonstrating a comparable performance to a benchtop commercial instrument. Paired with a smartphone for results visualization and geo-localization, this portable diagnostic platform with secure cloud connectivity will enable real-time case identification and epidemiological surveillance.

Nucleic acid amplification tests on respiratory samples for the diagnosis of coronavirus infections: a systematic review and meta-analysis

“… 43 Altogether the 51 studies analyzed 18,923 persons, of them 10,181 persons (22 studies) before COVID-19 and 8,742 persons for SARS-CoV-2 (29 studies). 7 , 9 , 13 , 14 , 17 , 18 , 19 , 22 , 23 , 25 , 26 , 27 , 34 , 35 , 36 , 38 , 39 , 41 , 43 , 45 , 46 , 47 , 50 , 51 , 53 , 54 , 55 , 56 The studies evaluated mostly patients with non-specific ILI or suspected COVID-19, the latter including both upper ARTI and pneumonia. Five studies included only children <2 years 16 , 21 , 31 , 32 , 48 , 25 studies included a mixed age range and 21 did not address patients' age.…”

Diagnosis of SARS-CoV-2 infection with LamPORE, a high-throughput platform combining loop-mediated isothermal amplification and nanopore sequencing

“…The copyright holder for this this version posted September 25, 2020. ; https://doi.org/10.1101/2020.09.18.20195370 doi: medRxiv preprint Kingdom testing program more than seven months after the SARS-CoV-2 pandemic was declared a public health emergency of international concern by the WHO. Further expansion of testing to include screening of asymptomatic individuals, which may be needed to prevent SARS-CoV-2 circulation, would require a significant further increase in testing capacity 1,2 . In the UK, clinical laboratories have struggled to expand conventional RT-PCR workflows to meet the demand for SARS-CoV-2 testing, and many have explored alternative methods that would be more scalable or allow near-patient use 3,4 . At the Oxford University Hospitals NHS Foundation Trust (OUH) and Sheffield Teaching Hospitals NHS Foundation Trust (STH), we evaluated LamPORE, a novel diagnostic platform for SARS-CoV-2 that combines loopmediated isothermal amplification (LAMP) with nanopore sequencing 5 .…”

LAMP Diagnostics at the Point-of-Care: Emerging Trends and Perspectives for the Developer Community

“…Arguably, LAMP is most valuable at the POC because it is fast, simple, and robust; and there are a few examples in the literature of POC platforms that incorporate RT-LAMP to detect SARS-CoV-2 (Table 2). Rodriguez-Manzano et al designed a lab on a chip platform capable of detecting 8 copies/µL of SARS-CoV-2 [132]. Although this handheld platform has a 91% sensitivity and 100% specificity when compared to RT-qPCR, RNA extraction is necessary prior to RT-LAMP [132].…”

“…Rodriguez-Manzano et al designed a lab on a chip platform capable of detecting 8 copies/µL of SARS-CoV-2 [132]. Although this handheld platform has a 91% sensitivity and 100% specificity when compared to RT-qPCR, RNA extraction is necessary prior to RT-LAMP [132]. Garneret et al describe their POC platform that utilizes membranes for RNA extraction and amplification [133].…”