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SARS-CoV-2: Comparative analysis of different RNA extraction methods.
Ambrosi, Cecilia; Prezioso, Carla; Checconi, Paola; Scribano, Daniela; Sarshar, Meysam; Capannari, Maurizio; Tomino, Carlo; Fini, Massimo; Garaci, Enrico; Palamara, Anna Teresa; De Chiara, Giovanna; Limongi, Dolores.
  • Ambrosi C; San Raffaele Roma Open University, 00166 Rome, Italy; IRCCS San Raffaele Pisana, 00166 Rome, Italy. Electronic address: cecilia.ambrosi@uniroma5.it.
  • Prezioso C; IRCCS San Raffaele Pisana, 00166 Rome, Italy; Department of Public Health and Infectious Diseases, Sapienza University of Rome, 00185 Rome, Italy.
  • Checconi P; San Raffaele Roma Open University, 00166 Rome, Italy; IRCCS San Raffaele Pisana, 00166 Rome, Italy.
  • Scribano D; Department of Public Health and Infectious Diseases, Sapienza University of Rome, 00185 Rome, Italy; Dani Di Giò Foundation-Onlus, 00193 Rome, Italy.
  • Sarshar M; Department of Public Health and Infectious Diseases, Sapienza University of Rome, Laboratory Affiliated to Institute Pasteur Italia- Cenci Bolognetti Foundation, 00185 Rome, Italy; Research Laboratories, Bambino Gesù Children's Hospital, IRCCS, 00146 Rome, Italy.
  • Capannari M; Elettrobiochimica s.r.l., 00159 Rome, Italy.
  • Tomino C; San Raffaele Roma Open University, 00166 Rome, Italy; IRCCS San Raffaele Pisana, 00166 Rome, Italy.
  • Fini M; IRCCS San Raffaele Pisana, 00166 Rome, Italy.
  • Garaci E; San Raffaele Roma Open University, 00166 Rome, Italy.
  • Palamara AT; IRCCS San Raffaele Pisana, 00166 Rome, Italy; Department of Public Health and Infectious Diseases, Sapienza University of Rome, Laboratory Affiliated to Institute Pasteur Italia- Cenci Bolognetti Foundation, 00185 Rome, Italy.
  • De Chiara G; Institute of Translational Pharmacology, National Research Council (CNR), Rome, Italy.
  • Limongi D; San Raffaele Roma Open University, 00166 Rome, Italy; IRCCS San Raffaele Pisana, 00166 Rome, Italy.
J Virol Methods ; 287: 114008, 2021 01.
Article in English | MEDLINE | ID: covidwho-907063
ABSTRACT
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiologic agent of the COVID-19 pandemic. Although other diagnostic methods have been introduced, detection of viral genes on oro- and nasopharyngeal swabs by reverse-transcription real time-PCR (rRT-PCR) assays is still the gold standard. Efficient viral RNA extraction is a prerequisite for downstream performance of rRT-PCR assays. Currently, several automatic methods that include RNA extraction are available. However, due to the growing demand, a shortage in kit supplies could be experienced in several labs. For these reasons, the use of different commercial or in-house protocols for RNA extraction may increase the possibility to analyze high number of samples. Herein, we compared the efficiency of RNA extraction of three different commercial kits and an in-house extraction protocol using synthetic ssRNA standards of SARS-CoV-2 as well as in oro-nasopharyngeal swabs from six COVID-19-positive patients. It was concluded that tested commercial kits can be used with some modifications for the detection of the SARS-CoV-2 genome by rRT-PCR approaches, although with some differences in RNA yields. Conversely, EXTRAzol reagent was the less efficient due to the phase separation principle at the basis of RNA extraction. Overall, this study offers alternative suitable methods to manually extract RNA that can be taken into account for SARS-CoV-2 detection.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Etiology study Limits: Humans Language: English Journal: J Virol Methods Year: 2021 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Etiology study Limits: Humans Language: English Journal: J Virol Methods Year: 2021 Document Type: Article