Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction.
Front Cell Infect Microbiol
; 11: 717068, 2021.
Artigo
em Inglês
| MEDLINE | ID: covidwho-1528814
ABSTRACT
This study aimed to detect the SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positive and 50 negative samples were divided into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at different temperatures (25°C, 60°C, and 98°C) and stored at 4°C. Group II was directly subjected to 3-step-heat treatment without PK exposure and stored at 4°C. And group III was set-up as standard group; it was processed using Qiagen's column based QIAamp Nucleic Acid kit and the obtained nucleic acids were stored at 4°C. These stored samples were used as a template to execute real-time polymerase chain reaction, and results were noted. Group I demonstrated 96% and 88% sensitivity for N and ORF1ab genes respectively, whereas group II demonstrated 78% and 60% when compared to the results of standard group III. Overall group I showed better results than group II when compared to group III. Thus, in situations where gold-standard reagents are not available, PK exposure and heat treatment can be employed to carry out molecular detection of SARS-CoV2 viral component.
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Texto completo:
Disponível
Coleções:
Bases de dados internacionais
Base de dados:
MEDLINE
Assunto principal:
RNA Viral
/
COVID-19
Tipo de estudo:
Estudo diagnóstico
/
Estudo experimental
/
Ensaios controlados aleatorizados
Limite:
Humanos
Idioma:
Inglês
Revista:
Front Cell Infect Microbiol
Ano de publicação:
2021
Tipo de documento:
Artigo
País de afiliação:
Fcimb.2021.717068
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