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Evaluation of recombinase-based isothermal amplification assays for point-of-need detection of SARS-CoV-2 in resource-limited settings.
Ghosh, Prakash; Chowdhury, Rajashree; Hossain, Mohammad Enayet; Hossain, Faria; Miah, Mojnu; Rashid, Md Utba; Baker, James; Rahman, Mohammed Ziaur; Rahman, Mustafizur; Ma, Xuejun; Duthie, Malcolm S; Wahed, Ahmed Abd El; Mondal, Dinesh.
  • Ghosh P; Emerging Infections and Parasitology Laboratory, NCSD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Chowdhury R; Emerging Infections and Parasitology Laboratory, NCSD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Hossain ME; Virology Laboratory, IDD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Hossain F; Emerging Infections and Parasitology Laboratory, NCSD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Miah M; Virology Laboratory, IDD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Rashid MU; Emerging Infections and Parasitology Laboratory, NCSD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Baker J; Laboratory Science and Services Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Rahman MZ; Virology Laboratory, IDD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Rahman M; Virology Laboratory, IDD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh.
  • Ma X; Chinese Center for Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, Beijing, 102206, China.
  • Duthie MS; HDT Bio Corp, Suite 280, 1616 Eastlake Ave E, Seattle, WA, 98102, USA.
  • Wahed AAE; Institute of Animal Hygiene and Veterinary Public Health, University of Leipzig, An den Tierkliniken 43, D-04103, Leipzig, Germany. Electronic address: ahmed.abd_el_wahed@uni-leipzig.de.
  • Mondal D; Emerging Infections and Parasitology Laboratory, NCSD, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh; Laboratory Science and Services Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, 1212, Bangladesh. E
Int J Infect Dis ; 114: 105-111, 2022 Jan.
Article in English | MEDLINE | ID: covidwho-1587637
ABSTRACT

OBJECTIVES:

The democratization of diagnostics is one of the key challenges towards containing the transmission of coronavirus disease 2019 (COVID-19) around the globe. The operational complexities of existing PCR-based methods, including sample transfer to advanced central laboratories with expensive equipment, limit their use in resource-limited settings. However, with the advent of isothermal technologies, the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is possible at decentralized facilities.

METHODS:

In this study, two recombinase-based isothermal techniques, reverse transcription recombinase polymerase amplification (RT-RPA) and reverse transcription recombinase-aided amplification (RT-RAA), were evaluated for the detection of SARS-CoV-2 in clinical samples. A total of 76 real-time reverse transcription PCR (real-time RT-PCR) confirmed COVID-19 cases and 100 negative controls were evaluated to determine the diagnostic performance of the isothermal methods.

RESULTS:

This investigation revealed equally promising diagnostic accuracy of the two methods, with a sensitivity of 76.32% (95% confidence interval 65.18-85.32%) when the target genes were RdRP and ORF1ab for RT-RPA and RT-RAA, respectively; the combination of N and RdRP in RT-RPA augmented the accuracy of the assay at a sensitivity of 85.53% (95% confidence interval 75.58-92.55%). Furthermore, high specificity was observed for each of the methods, ranging from 94.00% to 98.00% (95% confidence interval 87.40-9.76%).

CONCLUSIONS:

Considering the diagnostic accuracies, both RT-RPA and RT-RAA appear to be suitable assays for point-of-need deployment for the detection of the pathogen, understanding its epidemiology, case management, and curbing transmission.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Experimental Studies / Observational study / Prognostic study Limits: Humans Language: English Journal: Int J Infect Dis Journal subject: Communicable Diseases Year: 2022 Document Type: Article Affiliation country: J.ijid.2021.11.007

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Experimental Studies / Observational study / Prognostic study Limits: Humans Language: English Journal: Int J Infect Dis Journal subject: Communicable Diseases Year: 2022 Document Type: Article Affiliation country: J.ijid.2021.11.007