Development and Validation of a Rapid, Single-Step Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) System Potentially to Be Used for Reliable and High-Throughput Screening of COVID-19.
Front Cell Infect Microbiol
; 10: 331, 2020.
Artigo
em Inglês
| MEDLINE | ID: covidwho-634362
Preprint
Este artigo de periódico científico é provavelmente baseado em um preprint previamente disponível, por meio do reconhecimento de similaridade realizado por uma máquina. A confirmação humana ainda está pendente.
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Este artigo de periódico científico é provavelmente baseado em um preprint previamente disponível, por meio do reconhecimento de similaridade realizado por uma máquina. A confirmação humana ainda está pendente.
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ABSTRACT
Objectives:
Development and validation of a single-step and accurate reverse transcriptase loop-mediated isothermal amplification technique (RT-LAMP) for rapid identification of SARS-CoV-2 relative to commercial quantitative reverse transcriptase real-time PCR (qRT-PCR) assays to allow prompt initiation of proper medical care and containment of virus spread.Methods:
Primers showing optimal in-silico features were subjected to analytical sensitivity and specificity to assess the limit of detection (LOD) and cross-reaction with closely- and distantly-related viral species, and clinically prominent bacterial and fungal species. In order to evaluate the clinical utility, our RT-LAMP was subjected to a large number of clinical samples, including 213 negative and 47 positive patients, relative to two commercial quantitative RT-PCR assays.Results:
The analytical specificity and sensitivity of our assay was 100% and 500 copies/ml when serial dilution was performed in both water and sputum. Subjecting our RT-LAMP assay to clinical samples showed a high degree of specificity (99.5%), sensitivity (91.4%), positive predictive value (97.7%), and negative predictive value (98.1%) when used relative to qRT-PCR. Our RT-LAMP assay was two times faster than qRT-PCR and is storable at room temperature. A suspected case that later became positive tested positive using both our RT-LAMP and the two qRT-PCR assays, which shows the capability of our assay for screening purposes.Conclusions:
We present a rapid RT-LAMP assay that could extend the capacity of laboratories to process two times more clinical samples relative to qRT-PCR and potentially could be used for high-throughput screening purposes when demand is increasing at critical situations.Palavras-chave
Texto completo:
Disponível
Coleções:
Bases de dados internacionais
Base de dados:
MEDLINE
Assunto principal:
Pneumonia Viral
/
Programas de Rastreamento
/
Infecções por Coronavirus
/
Técnicas de Amplificação de Ácido Nucleico
/
Betacoronavirus
Tipo de estudo:
Estudo diagnóstico
/
Estudo experimental
/
Estudo prognóstico
/
Ensaios controlados aleatorizados
Limite:
Humanos
Idioma:
Inglês
Revista:
Front Cell Infect Microbiol
Ano de publicação:
2020
Tipo de documento:
Artigo
País de afiliação:
Fcimb.2020.00331
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