Cost-effective in-house COVID-19 reverse transcription-polymerase chain reaction testing with yeast-derived Taq polymerase

• Published in: Annals of thoracic medicine Vol. 19; no. 2; pp. 165 - 171
• Main Authors: Zhra, Mahmoud, Al Saud, Aljohara, Alzayer, Maha, Okdah, Liliane, Tamim, Hani, Fakhoury, Hana M A, Aljada, Ahmad
• Format: Journal Article
• Language: English
• Published: India Medknow Publications & Media Pvt. Ltd 01.04.2024; Wolters Kluwer - Medknow; Wolters Kluwer Medknow Publications
• Subjects: COVID-19; 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; Genes; Original; Pandemics; Polymerase chain reaction; RNA polymerase; Yeast; COVID-19; SARS-CoV-2; reverse transcription-polymerase chain reaction specificity; DNA contamination; recombinant yeast Taq polymerase; reverse transcription-polymerase chain reaction sensitivity; Taq polymerase
• ISSN: 1817-1737; 1998-3557
• DOI: 10.4103/atm.atm_180_23

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 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. This study investigates the production of recombinant 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 and genes, with human serving as an internal control. The unpurified yeast polymerase demonstrates sensitivity comparable to commercially purified bacterial polymerase and unpurified bacterial counterparts in detecting the and genes. It exhibits the highest specificity, with 100% accuracy, for the gene. The specificity for the gene closely aligns with that of commercially purified bacterial polymerase and unpurified bacterial polymerase. The use of unpurified recombinant yeast 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.