Academic Journal
Advancing COVID-19 diagnostics:rapid detection of intact SARS-CoV-2 using viability RT-PCR assay
| العنوان: | Advancing COVID-19 diagnostics:rapid detection of intact SARS-CoV-2 using viability RT-PCR assay |
|---|---|
| المؤلفون: | Veugen, Judith M.J., Schoenmakers, Tom, van Loo, Inge H.M., Haagmans, Bart L., Leers, Mathie P.G., Lamers, Mart M., Lucchesi, Mayk, van Bussel, Bas C.T., van Mook, Walther N.K.A., Nuijts, Rudy M.M.A., Savelkoul, Paul H.M., Dickman, Mor M., Wolffs, Petra F.G. |
| المصدر: | Veugen , J M J , Schoenmakers , T , van Loo , I H M , Haagmans , B L , Leers , M P G , Lamers , M M , Lucchesi , M , van Bussel , B C T , van Mook , W N K A , Nuijts , R M M A , Savelkoul , P H M , Dickman , M M & Wolffs , P F G 2024 , ' Advancing COVID-19 diagnostics : rapid detection of intact SARS-CoV-2 using viability RT-PCR assay ' , Microbiology spectrum , vol. 12 , .... |
| سنة النشر: | 2024 |
| مصطلحات موضوعية: | /dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being, name=SDG 3 - Good Health and Well-being |
| الوصف: | Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19). Commonly used methods for both clinical diagnosis of SARS-CoV-2 infection and management of infected patients involve the detection of viral RNA, but the presence of infectious virus particles is unknown. Viability PCR (v-PCR) uses a photoreactive dye to bind non-infectious RNA, ideally resulting in the detection of RNA only from intact virions. This study aimed to develop and validate a rapid v-PCR assay for distinguishing intact and compromised SARS-CoV-2. Propidium monoazide (PMAxx) was used as a photoreactive dye. Mixtures with decreasing percentages of intact SARS-CoV-2 (from 100% to 0%) were prepared from SARS-CoV-2 virus stock and a clinical sample. Each sample was divided into a PMAxx-treated part and a non-PMAxx-treated part. Reverse transcription-PCR (RT-PCR) using an in-house developed SARS-CoV-2 viability assay was then applied to both sample sets. The difference in intact SARS-CoV-2 was determined by subtracting the cycle threshold (Ct) value of the PMAxx-treated sample from the non-PMAxx-treated sample. Mixtures with decreasing concentrations of intact SARS-CoV-2 showed increasingly lower delta Ct values as the percentage of intact SARS-CoV-2 decreased, as expected. This relationship was observed in both high and low viral load samples prepared from cultured SARS-CoV-2 virus stock, as well as for a clinical sample prepared directly from a SARS-CoV-2 positive nasopharyngeal swab. In this study, a rapid v-PCR assay has been validated that can distinguish intact from compromised SARS-CoV-2. The presence of intact virus particles, as determined by v-PCR, may indicate SARS-CoV-2 infectiousness. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| DOI: | 10.1128/spectrum.00160-24 |
| الاتاحة: | https://pure.eur.nl/en/publications/6e50de9d-20cd-4f8f-90fb-eb6f8b44035b https://doi.org/10.1128/spectrum.00160-24 https://pure.eur.nl/ws/files/167534148/veugen-et-al-2024-advancing-covid-19-diagnostics-rapid-detection-of-intact-sars-cov-2-using-viability-rt-pcr-assay.pdf http://www.scopus.com/inward/record.url?scp=85203205803&partnerID=8YFLogxK |
| Rights: | info:eu-repo/semantics/openAccess |
| رقم الانضمام: | edsbas.5210ABD3 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1128/spectrum.00160-24 |
|---|