Point-of-care testing for COVID-19: a simple two-step molecular diagnostic development and validation during the SARS-CoV-2 pandemic.

Autor:	Yoshikawa AAG; Universidade Federal de Santa Catarina, Departamento de Biologia Celular, Embriologia e Genética, Florianópolis, SC, Brasil., Cardoso SF; Universidade Federal de Santa Catarina, Departamento de Biologia Celular, Embriologia e Genética, Florianópolis, SC, Brasil.; Secretaria de Saúde do Estado de Santa Catarina, Diretoria de Vigilância Epidemiológica, Florianópolis, SC, Brasil., Eslabão LB; Universidade Federal de Santa Catarina, Departamento de Microbiologia, Imunologia e Parasitologia, Florianópolis, SC, Brasil., Pinheiro IC; Universidade Federal de Santa Catarina, Departamento de Biologia Celular, Embriologia e Genética, Florianópolis, SC, Brasil., Valverde P; Secretaria Municipal de Saúde, Florianópolis, SC, Brasil., Caminha G; Laboratório Central de Saúde Pública de Santa Catarina, Florianópolis, SC, Brasil., Romero OB; Universidade Federal de Santa Catarina, Departamento de Microbiologia, Imunologia e Parasitologia, Florianópolis, SC, Brasil., Medeiros L; Instituto Federal de Educação, Ciência e Tecnologia de Santa Catarina, Florianópolis, SC, Brasil., Rona LDP; Universidade Federal de Santa Catarina, Departamento de Biologia Celular, Embriologia e Genética, Florianópolis, SC, Brasil.; Conselho Nacional de Desenvolvimento Científico e Tecnológico, Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Rio de Janeiro, RJ, Brasil., Pitaluga AN; Conselho Nacional de Desenvolvimento Científico e Tecnológico, Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Rio de Janeiro, RJ, Brasil.; Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.
Jazyk:	angličtina
Zdroj:	Memorias do Instituto Oswaldo Cruz [Mem Inst Oswaldo Cruz] 2024 Oct 04; Vol. 119, pp. e230236. Date of Electronic Publication: 2024 Oct 04 (Print Publication: 2024).
DOI:	10.1590/0074-02760230236
Abstrakt:	Background: During the coronavirus disease 19 (COVID-19) pandemic, diagnostic testing of the general population proved challenging due to limitations of the gold-standard diagnostic procedure using reverse transcription real-time polymerase chain reaction (RT-qPCR) for large-scale testing on the centralised model, especially in low-resource areas. Objectives: To address this, a point-of-care (PoC) diagnostic protocol for COVID-19 was developed, providing fast, reliable, and affordable testing, particularly for low-mid develop areas. Methods: The PoC diagnostic process combines a simple paper-based RNA extraction method housed within a 3D-printed plastic device with a colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay. Nasopharyngeal/oropharyngeal swabs (NOS) and saliva samples were tested between 2020 and 2021, with the assistance of Santa Catarina's State Health Secretary, Brazil. Findings: The developed diagnostic protocol showed a limit of detection of 9,900 copies and an overall diagnostic specificity of 98% for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from 1,348 clinical analysed samples. The diagnostic sensitivity was 95% for NOS samples, 85% for early morning saliva, and 69% for indiscriminate saliva. Main Conclusions: In conclusion, the developed device successfully extracted SARS-CoV-2 viral RNA from swabs and saliva clinical samples. When combined with colorimetric RT-LAMP, it provides results within 45 min using minimal resources, thus delivering a diagnostic kit protocol that is applicable in large-scale sampling.
Databáze:	MEDLINE
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