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Digital RT-PCR Chip method for detection of SARS-CoV-2 virus

The “gold standard” method for detection of SARS-CoV-2 is the real time reverse transcription-polymerase chain reaction, but due to pre-analytical and technical limitations, biological samples with low viral load are not sometimes detected. For this purpose a digital RT-PCR method on-chip was develo...

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Detalles Bibliográficos
Autores principales: Dioni, Laura, Orlandi, Annarosa, Uceda Renteria, Sara, Favero, Chiara, Solazzo, Giulia, Oggioni, Massimo, Bollati, Valentina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9383957/
https://www.ncbi.nlm.nih.gov/pubmed/35985558
http://dx.doi.org/10.1016/j.jim.2022.113339
Descripción
Sumario:The “gold standard” method for detection of SARS-CoV-2 is the real time reverse transcription-polymerase chain reaction, but due to pre-analytical and technical limitations, biological samples with low viral load are not sometimes detected. For this purpose a digital RT-PCR method on-chip was developed for detection of the SARS-CoV-2 virus, using two TaqMan™ Assays for quantification of the N Protein (Nucleocapsid) and the S Protein (Spike), and the QuantStudio™ 3D Digital PCR instrument. The method was applied to assess the nasopharyngeal swabs of asymptomatic subjects recruited in the UNICORN Study. The digital RT-PCR method is characterized by a higher sensitivity than the RT-qPCR method, even if performed with the same TaqMan™, and could be a promising tool for SARS-CoV-2 viral load quantification.