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Am J Clin Pathol . Asymptomatic Patient Testing After 10:1 Pooling Using the Xpert Xpress SARS-CoV-2 Assay

tetano

Editor, Senior Moderator
Am J Clin Pathol


. 2021 Jan 5;aqaa273.
doi: 10.1093/ajcp/aqaa273. Online ahead of print.
Asymptomatic Patient Testing After 10:1 Pooling Using the Xpert Xpress SARS-CoV-2 Assay


Gary W Procop[SUP] 1 [/SUP], Marion Tuohy[SUP] 1 [/SUP], Christine Ramsey[SUP] 1 [/SUP], Daniel D Rhoads[SUP] 1 [/SUP], Brian P Rubin[SUP] 1 [/SUP], Richard Figler[SUP] 2 [/SUP]



Affiliations

Abstract

Objectives: Pool testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) preserves testing resources at the risk of missing specimens through specimen dilution.
Methods: To determine whether SARS-CoV-2 specimens would be missed after 10:1 pooling, we identified 10 specimens with midrange (ie, 25-34 cycles) and 10 with late (ie, >34-45 cycles) crossing threshold (Ct) values and tested these both neat and after 10:1 pooling. Final test results and Ct changes were compared.
Results: Overall, 17 of 20 specimens that contained SARS-CoV-2 were detected after 10:1 pooling with the Xpert Xpress SARS-CoV-2 Assay (Cepheid), rendering an 85% positive percentage of agreement. All 10 of 10 specimens with an undiluted Ct in the mid-Ct range were detected after 10:1 pooling, in contrast to 7 of 10 with an undiluted Ct in the late-Ct range. The overall Ct difference between the neat testing and the 10:1 pool was 2.9 cycles for the N2 gene target and 3 cycles for the E gene target. The N2 gene reaction was more sensitive than the E gene reaction, detecting 16 of 20 positive specimens after 10:1 pooling compared with 9 of 20 specimens.
Conclusions: An 85% positive percentage of agreement was achieved, with only specimens with low viral loads being missed following 10:1 pooling. The average impact on both reverse transcription polymerase chain reactions within this assay was about 3 cycles.

Keywords: COVID-19; Coronavirus; Pooling; SARS-CoV-2; Xpert Xpress SARS-CoV-2 assay.
 
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