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Performance of Nucleic Acid Amplification Tests for Detection of Severe Acute Respiratory Syndrome Coronavirus 2 in Prospectively Pooled Specimens EMERGING INFECTIOUS DISEASES Wang, H., Hogan, C. A., Miller, J. A., Sahoo, M. K., Huang, C., Mfuh, K. O., Sibai, M., Zehnder, J., Hickey, B., Sinnott-Armstrong, N., Pinsky, B. A. 2021; 27 (1): 92–103

Abstract

Pooled nucleic acid amplification tests for severe acute respiratory syndrome coronavirus 2 could increase availability of testing at decreased cost. However, the effect of dilution on analytical sensitivity through sample pooling has not been well characterized. We tested 1,648 prospectively pooled specimens by using 3 nucleic acid amplification tests for severe acute respiratory syndrome coronavirus 2: a laboratory-developed real-time reverse transcription PCR targeting the envelope gene, and 2 commercially available Panther System assays targeting open reading frame 1ab. Positive percent agreement (PPA) of pooled versus individual testing ranged from 71.7% to 82.6% for pools of 8 and from 82.9% to 100.0% for pools of 4. We developed and validated an independent stochastic simulation model to estimate effects of dilution on PPA and efficiency of a 2-stage pooled real-time reverse transcription PCR testing algorithm. PPA was dependent on the proportion of tests with positive results, cycle threshold distribution, and assay limit of detection.

View details for DOI 10.3201/eid2701.203379

View details for Web of Science ID 000609135300011

View details for PubMedID 33183494

View details for PubMedCentralID PMC7774575