tetano
Editor, Senior Moderator
Sci Rep
. 2020 Oct 6;10(1):16608.
doi: 10.1038/s41598-020-73616-w.
A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
Aniela Wozniak[SUP] 1 [/SUP], Ariel Cerda[SUP] 2 [/SUP], Catalina Ibarra-Henr?quez[SUP] 2 [/SUP], Valentina Sebastian[SUP] 3 [/SUP], Grace Armijo[SUP] 2 [/SUP], Liliana Lamig[SUP] 2 [/SUP], Carolina Miranda[SUP] 3 [/SUP], Marcela Lagos[SUP] 1 [/SUP], Sandra Solari[SUP] 1 [/SUP], Ana Mar?a Guzm?n[SUP] 1 [/SUP], Teresa Quiroga[SUP] 1 [/SUP], Susan Hitschfeld[SUP] 2 [/SUP], Eleodoro Riveras[SUP] 2 [/SUP], Marcela Ferr?s[SUP] 1 4 [/SUP], Rodrigo A Guti?rrez[SUP] 5 [/SUP], Patricia Garc?a[SUP] 6 [/SUP]
Affiliations
Abstract
The technique RT-qPCR for viral RNA detection is the current worldwide strategy used for early detection of the novel coronavirus SARS-CoV-2. RNA extraction is a key pre-analytical step in RT-qPCR, often achieved using commercial kits. However, the magnitude of the COVID-19 pandemic is causing disruptions to the global supply chains used by many diagnostic laboratories to procure the commercial kits required for RNA extraction. Shortage in these essential reagents is even more acute in developing countries with no means to produce kits locally. We sought to find an alternative procedure to replace commercial kits using common reagents found in molecular biology laboratories. Here we report a method for RNA extraction that takes about 40 min to complete ten samples, and is not more laborious than current commercial RNA extraction kits. We demonstrate that this method can be used to process nasopharyngeal swab samples and yields RT-qPCR results comparable to those obtained with commercial kits. Most importantly, this procedure can be easily implemented in any molecular diagnostic laboratory. Frequent testing is crucial for individual patient management as well as for public health decision making in this pandemic. Implementation of this method could maintain crucial testing going despite commercial kit shortages.
. 2020 Oct 6;10(1):16608.
doi: 10.1038/s41598-020-73616-w.
A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
Aniela Wozniak[SUP] 1 [/SUP], Ariel Cerda[SUP] 2 [/SUP], Catalina Ibarra-Henr?quez[SUP] 2 [/SUP], Valentina Sebastian[SUP] 3 [/SUP], Grace Armijo[SUP] 2 [/SUP], Liliana Lamig[SUP] 2 [/SUP], Carolina Miranda[SUP] 3 [/SUP], Marcela Lagos[SUP] 1 [/SUP], Sandra Solari[SUP] 1 [/SUP], Ana Mar?a Guzm?n[SUP] 1 [/SUP], Teresa Quiroga[SUP] 1 [/SUP], Susan Hitschfeld[SUP] 2 [/SUP], Eleodoro Riveras[SUP] 2 [/SUP], Marcela Ferr?s[SUP] 1 4 [/SUP], Rodrigo A Guti?rrez[SUP] 5 [/SUP], Patricia Garc?a[SUP] 6 [/SUP]
Affiliations
- PMID: 33024174
- DOI: 10.1038/s41598-020-73616-w
Abstract
The technique RT-qPCR for viral RNA detection is the current worldwide strategy used for early detection of the novel coronavirus SARS-CoV-2. RNA extraction is a key pre-analytical step in RT-qPCR, often achieved using commercial kits. However, the magnitude of the COVID-19 pandemic is causing disruptions to the global supply chains used by many diagnostic laboratories to procure the commercial kits required for RNA extraction. Shortage in these essential reagents is even more acute in developing countries with no means to produce kits locally. We sought to find an alternative procedure to replace commercial kits using common reagents found in molecular biology laboratories. Here we report a method for RNA extraction that takes about 40 min to complete ten samples, and is not more laborious than current commercial RNA extraction kits. We demonstrate that this method can be used to process nasopharyngeal swab samples and yields RT-qPCR results comparable to those obtained with commercial kits. Most importantly, this procedure can be easily implemented in any molecular diagnostic laboratory. Frequent testing is crucial for individual patient management as well as for public health decision making in this pandemic. Implementation of this method could maintain crucial testing going despite commercial kit shortages.