Agile design and development of a high throughput cobas SARS-CoV-2 RT-PCR diagnostic test

被引：1
|
作者：
Manohar, Chitra [1 ]
Sun, Jingtao [1 ]
Schlag, Peter [1 ]
Santini, Chris [1 ]
Fontecha, Marcel [1 ]
Lotscher, Pirmin [2 ]
Bier, Carolin [2 ]
Goepfert, Kristina [2 ]
Duncan, Dana [1 ]
Spier, Gene [1 ]
Jarem, Daniel [1 ]
Kosarikov, Dmitriy [1 ]
机构：
[1] Roche Mol Syst Inc, Pleasanton, CA 94588 USA
[2] Roche Diagnost Int AG, Rotkreuz, Switzerland
来源：
HELIYON | 2022年 / 8卷 / 09期
基金：
欧盟地平线“2020”;
关键词：
SARS-CoV-2; COVID-19; Pandemic; RT-PCR; Diagnostic testing; Molecular assay design;
D O I：
10.1016/j.heliyon.2022.e10591
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
Diagnostic testing is essential for management of the COVID-19 pandemic. An agile assay design methodology, optimized for the cobas (R) 6800/8800 system, was used to develop a dual-target, qualitative SARS-CoV-2 RT-PCR test using commercially available reagents and existing sample processing and thermocycling profiles. The limit of detection was 30-52 copies/mL for USA-WA1/2020. Assay sensitivity was confirmed for SARS-CoV-2 variants Alpha, Beta, Gamma, Delta and Kappa. The coefficients of variation of the cycle threshold number (Ct) were between 1.1 and 2.2%. There was no difference in Ct using nasopharyngeal compared to oropharyngeal swabs in universal transport medium (UTM). A small increase in Ct was observed with specimens collected in cobas PCR medium compared to UTM. In silico analysis indicated that the dual-target test is capable of detecting all >1,800,000 SARS-CoV-2 sequences in the GISAID database. Our agile assay design approach facilitated rapid development and deployment of this SARS-CoV-2 RT-PCR test.
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页数：8
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