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The Production of Standardized Samples with Known Concentrations for Severe Acute Respiratory Syndrome Coronavirus 2 RT-qPCR Testing Validation for Developing Countries in the Period of the Pandemic Era
BioMed Research International Số , năm 2021 (Tập 2021, trang -)
ISSN: 23146133
ISSN: 23146133
DOI: 10.1155/2021/5516344
Tài liệu thuộc danh mục:
Article
English
Từ khóa: COVID-19; Developing Countries; Humans; Molecular Diagnostic Techniques; Pandemics; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral; SARS-CoV-2; Validation Studies as Topic; virus DNA; virus RNA; virus RNA; Article; biosafety level 3; coronavirus disease 2019; COVID-19 nucleic acid testing; cycle threshold value; developing country; diagnostic accuracy; dilution; DNA sequence; DNA synthesis; in vitro study; limit of detection; measurement precision; measurement repeatability; nonhuman; pandemic; practice guideline; real time reverse transcription polymerase chain reaction; RNA extraction; RNA synthesis; sensitivity and specificity; Severe acute respiratory syndrome coronavirus 2; standardization; validation process; virus culture; virus inactivation; diagnosis; epidemiology; genetics; human; isolation and purification; molecular diagnosis; pandemic; procedures; reverse transcription polymerase chain reaction; validation study; virology
Tóm tắt tiếng anh
Background. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic of pneumonia spreading around the world, leading to serious threats to public health and attracting enormous attention. There is an urgent need for sensitive diagnostic testing implementation to control and manage SARS-CoV-2 in public health laboratories. The quantitative reverse transcription PCR (RT-qPCR) assay is the gold standard method, but the sensitivity and specificity of SARS-CoV-2 testing are dependent on a number of factors. Methods. We synthesized RNA based on the genes published to estimate the concentration of inactivated virus samples in a biosafety level 3 laboratory. The limit of detection (LOD), linearity, accuracy, and precision were evaluated according to the bioanalytical method validation guidelines. Results. We found that the LOD reached around 3 copies/reaction. Furthermore, intra-assay precision, accuracy, and linearity met the accepted criterion with an RSD for copies of less than 25%, and linear regression met the accepted R2 of 0.98. Conclusions. We suggest that synthesized RNA based on the database of the NCBI gene bank for estimating the concentration of inactivated virus samples provides a potential opportunity for reliable testing to diagnose coronavirus disease 2019 (COVID-19) as well as limit the spread of the disease. This method may be relatively quick and inexpensive, and it may be useful for developing countries during the pandemic era. In the long term, it is also applicable for evaluation, verification, validation, and external quality assessment. 2021 Hoang Quoc Cuong et al.