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
| Autor: | Tran Ton, Tran Cat Dong, Hoang Quoc Cuong, Huynh Thi Kim Loan, Hoang Thuy Linh, Nguyen Trung Hieu, Nguyen Thi Thanh Thao, Phan Trong Lan, Cao Minh Thang, Nguyen Duc Hai, Do Thi Hong Tuoi, Vu Thanh Thao, Nguyen Duc Tuan, Nguyen Thanh Long, Nguyen Hoang Anh |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Article Subject Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) 030106 microbiology Developing country Disease Validation Studies as Topic General Biochemistry Genetics and Molecular Biology Virus 03 medical and health sciences Biosafety level External quality assessment Pandemic Humans Medicine Developing Countries Pandemics General Immunology and Microbiology Reverse Transcriptase Polymerase Chain Reaction SARS-CoV-2 business.industry COVID-19 General Medicine Gold standard (test) Virology 030104 developmental biology Molecular Diagnostic Techniques RNA Viral business Research Article |
| Zdroj: | BioMed Research International, Vol 2021 (2021) BioMed Research International |
| ISSN: | 2314-6141 2314-6133 |
| Popis: | 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 R 2 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. |
| Databáze: | OpenAIRE |
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