Cost-effective in-house COVID-19 reverse transcription-polymerase chain reaction testing with yeast-derived Taq polymerase
| Autor: | Mahmoud Zhra, Aljohara Al Saud, Maha Alzayer, Liliane Okdah, Hani Tamim, Hana M. A Fakhoury, Ahmad Aljada |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2024 |
| Předmět: |
covid-19
dna contamination recombinant yeast taq polymerase reverse transcription-polymerase chain reaction sensitivity reverse transcription-polymerase chain reaction specificity sars-cov-2 taq polymerase Diseases of the circulatory (Cardiovascular) system RC666-701 Diseases of the respiratory system RC705-779 |
| Zdroj: | Annals of Thoracic Medicine, Vol 19, Iss 2, Pp 165-171 (2024) |
| Druh dokumentu: | article |
| ISSN: | 1817-1737 |
| DOI: | 10.4103/atm.atm_180_23 |
| Popis: | BACKGROUND: Despite the decline of the COVID-19 pandemic, there continues to be a persistent requirement for reliable testing methods that can be adapted to future outbreaks and areas with limited resources. While the standard approach of using reverse transcription-polymerase chain reaction (RT-PCR) with Taq polymerase is effective, it faces challenges such as limited access to high-quality enzymes and the presence of bacterial DNA contamination in commercial kits, which can impact the accuracy of test results. METHODS: This study investigates the production of recombinant Taq polymerase in yeast cells and assesses its crude lysate in a multiplex RT-PCR assay for detecting the SARS-CoV-2 RNA-dependent RNA polymerase (RdRP) and N genes, with human Ribonuclease P serving as an internal control. RESULTS: The unpurified yeast Taq polymerase demonstrates sensitivity comparable to commercially purified bacterial Taq polymerase and unpurified bacterial counterparts in detecting the RdRP and N genes. It exhibits the highest specificity, with 100% accuracy, for the N gene. The specificity for the RdRP gene closely aligns with that of commercially purified bacterial Taq polymerase and unpurified bacterial Taq polymerase. CONCLUSIONS: The use of unpurified recombinant yeast Taq polymerase shows promise as a cost-effective approach for conducting in-house COVID-19 RT-PCR testing. By eliminating the need for chromatography purification steps, the production of RT-PCR kits can be streamlined, potentially improving accessibility and scalability, especially in resource-limited settings and future pandemics. |
| Databáze: | Directory of Open Access Journals |
| Externí odkaz: |
|