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Unlocking SARS-CoV-2 detection in low- and middle-income countries

Alcántara, Roberto ; Peñaranda, Katherin ; Mendoza-Rojas, Gabriel ; Nakamoto, Jose A LU orcid ; Martins-Luna, Johanna ; Del Valle-Mendoza, Juana ; Adaui, Vanessa and Milón, Pohl (2021) In Cell reports methods 1(7).
Abstract

Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes and nucleic acids can supply over 50,000 tests per production batch. The optimized one-step RT-PCR coupled to CRISPR-Cas12a-mediated detection showed a limit of detection of 102 ge/μL in a turnaround time of 2 h. The clinical validation indicated an overall sensitivity of 80%-88%, while for middle and high viral load samples (Cq ≤ 31) the... (More)

Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes and nucleic acids can supply over 50,000 tests per production batch. The optimized one-step RT-PCR coupled to CRISPR-Cas12a-mediated detection showed a limit of detection of 102 ge/μL in a turnaround time of 2 h. The clinical validation indicated an overall sensitivity of 80%-88%, while for middle and high viral load samples (Cq ≤ 31) the sensitivity was 92%-100%. The specificity was 96%-100% regardless of viral load. Furthermore, we show that the toolkit can be used with the mobile laboratory Bento Lab, potentially enabling LMICs to implement detection services in unattended remote regions.

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author
; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
in
Cell reports methods
volume
1
issue
7
article number
100093
publisher
Cell Press
external identifiers
  • scopus:85118504823
  • pmid:34697612
ISSN
2667-2375
DOI
10.1016/j.crmeth.2021.100093
language
English
LU publication?
no
additional info
© 2021 The Authors.
id
a3f6fa7b-4c25-4a02-9436-76721f28d729
date added to LUP
2022-10-13 10:20:59
date last changed
2024-04-19 19:53:34
@article{a3f6fa7b-4c25-4a02-9436-76721f28d729,
  abstract     = {{<p>Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes and nucleic acids can supply over 50,000 tests per production batch. The optimized one-step RT-PCR coupled to CRISPR-Cas12a-mediated detection showed a limit of detection of 102 ge/μL in a turnaround time of 2 h. The clinical validation indicated an overall sensitivity of 80%-88%, while for middle and high viral load samples (Cq ≤ 31) the sensitivity was 92%-100%. The specificity was 96%-100% regardless of viral load. Furthermore, we show that the toolkit can be used with the mobile laboratory Bento Lab, potentially enabling LMICs to implement detection services in unattended remote regions.</p>}},
  author       = {{Alcántara, Roberto and Peñaranda, Katherin and Mendoza-Rojas, Gabriel and Nakamoto, Jose A and Martins-Luna, Johanna and Del Valle-Mendoza, Juana and Adaui, Vanessa and Milón, Pohl}},
  issn         = {{2667-2375}},
  language     = {{eng}},
  month        = {{11}},
  number       = {{7}},
  publisher    = {{Cell Press}},
  series       = {{Cell reports methods}},
  title        = {{Unlocking SARS-CoV-2 detection in low- and middle-income countries}},
  url          = {{http://dx.doi.org/10.1016/j.crmeth.2021.100093}},
  doi          = {{10.1016/j.crmeth.2021.100093}},
  volume       = {{1}},
  year         = {{2021}},
}