tetano
Editor, Senior Moderator
J Mol Diagn
. 2021 Mar 8;S1525-1578(21)00063-5.
doi: 10.1016/j.jmoldx.2021.02.009. Online ahead of print.
Microchip RT-PCR Detection of Nasopharyngeal SARS-CoV-2 Samples
Razvan Cojocaru[SUP] 1 [/SUP], Iqra Yaseen[SUP] 1 [/SUP], Peter J Unrau[SUP] 2 [/SUP], Christopher F Lowe[SUP] 3 [/SUP], Gordon Ritchie[SUP] 3 [/SUP], Marc G Romney[SUP] 3 [/SUP], Don D Sin[SUP] 4 [/SUP], Sikander Gill[SUP] 5 [/SUP], Maxim Slyadnev[SUP] 5 [/SUP]
Affiliations
Abstract
Fast, accurate and reliable diagnostic tests are critical for controlling the spread of the 2019 Coronavirus disease associated with SARS-CoV-2 infection. The current gold standard for testing is real time PCR (RT-PCR), however, during the current pandemic, supplies of testing kits and reagents have been limited. We report the validation of a rapid (30 min), user-friendly and accurate microchip RT-PCR assay for detection of SARS-CoV-2 from nasopharyngeal swab RNA extracts. Microchips pre-loaded with COVID-19 primers and probes for the N gene accommodate 1.2 ?l reaction volumes, lowering the required reagents by 10-fold compared to tube-based RT-PCR. We validated our assay using contrived reference samples and 21 clinical samples from patients in Canada, determining a limit of detection of 1 copy per reaction. The microchip RT-PCR provides a significantly lower resource alternative to the CDC approved real-time RT-PCR assays with comparable sensitivity, showing 100% positive and negative predictive agreement of clinical samples.
. 2021 Mar 8;S1525-1578(21)00063-5.
doi: 10.1016/j.jmoldx.2021.02.009. Online ahead of print.
Microchip RT-PCR Detection of Nasopharyngeal SARS-CoV-2 Samples
Razvan Cojocaru[SUP] 1 [/SUP], Iqra Yaseen[SUP] 1 [/SUP], Peter J Unrau[SUP] 2 [/SUP], Christopher F Lowe[SUP] 3 [/SUP], Gordon Ritchie[SUP] 3 [/SUP], Marc G Romney[SUP] 3 [/SUP], Don D Sin[SUP] 4 [/SUP], Sikander Gill[SUP] 5 [/SUP], Maxim Slyadnev[SUP] 5 [/SUP]
Affiliations
- PMID: 33706009
- DOI: 10.1016/j.jmoldx.2021.02.009
Abstract
Fast, accurate and reliable diagnostic tests are critical for controlling the spread of the 2019 Coronavirus disease associated with SARS-CoV-2 infection. The current gold standard for testing is real time PCR (RT-PCR), however, during the current pandemic, supplies of testing kits and reagents have been limited. We report the validation of a rapid (30 min), user-friendly and accurate microchip RT-PCR assay for detection of SARS-CoV-2 from nasopharyngeal swab RNA extracts. Microchips pre-loaded with COVID-19 primers and probes for the N gene accommodate 1.2 ?l reaction volumes, lowering the required reagents by 10-fold compared to tube-based RT-PCR. We validated our assay using contrived reference samples and 21 clinical samples from patients in Canada, determining a limit of detection of 1 copy per reaction. The microchip RT-PCR provides a significantly lower resource alternative to the CDC approved real-time RT-PCR assays with comparable sensitivity, showing 100% positive and negative predictive agreement of clinical samples.