tetano
Editor, Senior Moderator
J Clin Microbiol
. 2020 Jul 29;JCM.01669-20.
doi: 10.1128/JCM.01669-20. Online ahead of print.
Performance characteristics of a high throughput automated transcription mediated amplification test for SARS-CoV-2 detection
Jimmykim Pham[SUP] 1 [/SUP], Sarah Meyer[SUP] 1 [/SUP], Catherine Nguyen[SUP] 1 [/SUP], Analee Williams[SUP] 1 [/SUP], Melissa Hunsicker[SUP] 1 [/SUP], Ian McHardy[SUP] 2 [/SUP], Inessa Gendlina[SUP] 3 [/SUP], D Yitzchak Goldstein[SUP] 3 [/SUP], Amy S Fox[SUP] 3 [/SUP], Angela Hudson[SUP] 1 [/SUP], Paul Darby[SUP] 1 [/SUP], Paul Hovey[SUP] 1 [/SUP], Jose Morales[SUP] 1 [/SUP], James Mitchell[SUP] 1 [/SUP], Karen Harrington[SUP] 1 [/SUP], Mehrdad Majlessi[SUP] 1 [/SUP], Joshua Moberly[SUP] 1 [/SUP], Ankur Shah[SUP] 1 [/SUP], Andrew Worlock[SUP] 1 [/SUP], Marion Walcher[SUP] 1 [/SUP], Barbara Eaton[SUP] 1 [/SUP], Damon Getman[SUP] 4 [/SUP], Craig Clark[SUP] 1 [/SUP]
Affiliations
Abstract
The COVID-19 pandemic caused by the new SARS-CoV-2 coronavirus has imposed severe challenges on laboratories in their effort to achieve sufficient diagnostic testing capability for identifying infected individuals. In this study we report the analytical and clinical performance characteristics of a new, high-throughput, fully automated nucleic acid amplification test system for the detection of SARS-CoV-2. The assay utilizes target capture, transcription mediated amplification, and acridinium ester-labeled probe chemistry on the automated Panther System to directly amplify and detect two separate target sequences in the ORF1ab region of the SARS-CoV-2 RNA genome. The probit 95% limit of detection of the assay was determined to be 0.004 TCID[SUB]50[/SUB]/ml using inactivated virus, and 25 c/ml using synthetic in vitro transcript RNA targets. Analytical sensitivity (100% detection) was confirmed to be 83 - 194 c/ml using three commercially available SARS-CoV-2 nucleic acid controls. No cross reactivity or interference was observed with testing six related human coronaviruses, as well as 24 other viral, fungal, and bacterial pathogens, at high titer. Clinical nasopharyngeal swab specimen testing (N=140) showed 100%, 98.7%, and 99.3% positive, negative, and overall agreement, respectively, with a validated reverse transcription PCR NAAT for SARS-CoV-2 RNA. These results provide validation evidence for a sensitive and specific method for pandemic-scale automated molecular diagnostic testing for SARS-CoV-2.
. 2020 Jul 29;JCM.01669-20.
doi: 10.1128/JCM.01669-20. Online ahead of print.
Performance characteristics of a high throughput automated transcription mediated amplification test for SARS-CoV-2 detection
Jimmykim Pham[SUP] 1 [/SUP], Sarah Meyer[SUP] 1 [/SUP], Catherine Nguyen[SUP] 1 [/SUP], Analee Williams[SUP] 1 [/SUP], Melissa Hunsicker[SUP] 1 [/SUP], Ian McHardy[SUP] 2 [/SUP], Inessa Gendlina[SUP] 3 [/SUP], D Yitzchak Goldstein[SUP] 3 [/SUP], Amy S Fox[SUP] 3 [/SUP], Angela Hudson[SUP] 1 [/SUP], Paul Darby[SUP] 1 [/SUP], Paul Hovey[SUP] 1 [/SUP], Jose Morales[SUP] 1 [/SUP], James Mitchell[SUP] 1 [/SUP], Karen Harrington[SUP] 1 [/SUP], Mehrdad Majlessi[SUP] 1 [/SUP], Joshua Moberly[SUP] 1 [/SUP], Ankur Shah[SUP] 1 [/SUP], Andrew Worlock[SUP] 1 [/SUP], Marion Walcher[SUP] 1 [/SUP], Barbara Eaton[SUP] 1 [/SUP], Damon Getman[SUP] 4 [/SUP], Craig Clark[SUP] 1 [/SUP]
Affiliations
- PMID: 32727828
- DOI: 10.1128/JCM.01669-20
Abstract
The COVID-19 pandemic caused by the new SARS-CoV-2 coronavirus has imposed severe challenges on laboratories in their effort to achieve sufficient diagnostic testing capability for identifying infected individuals. In this study we report the analytical and clinical performance characteristics of a new, high-throughput, fully automated nucleic acid amplification test system for the detection of SARS-CoV-2. The assay utilizes target capture, transcription mediated amplification, and acridinium ester-labeled probe chemistry on the automated Panther System to directly amplify and detect two separate target sequences in the ORF1ab region of the SARS-CoV-2 RNA genome. The probit 95% limit of detection of the assay was determined to be 0.004 TCID[SUB]50[/SUB]/ml using inactivated virus, and 25 c/ml using synthetic in vitro transcript RNA targets. Analytical sensitivity (100% detection) was confirmed to be 83 - 194 c/ml using three commercially available SARS-CoV-2 nucleic acid controls. No cross reactivity or interference was observed with testing six related human coronaviruses, as well as 24 other viral, fungal, and bacterial pathogens, at high titer. Clinical nasopharyngeal swab specimen testing (N=140) showed 100%, 98.7%, and 99.3% positive, negative, and overall agreement, respectively, with a validated reverse transcription PCR NAAT for SARS-CoV-2 RNA. These results provide validation evidence for a sensitive and specific method for pandemic-scale automated molecular diagnostic testing for SARS-CoV-2.