Population testing for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is necessary owing to the potential for viral transmission from asymptomatic cases, yet scarcity of reagents and equipment has increased the cost-prohibitive implementation of screening campaigns at institutions of higher education. Significant analytical sensitivities of nucleic acid amplification methods permit sample pooling to increase testing capacity. Statistical models compared optimal testing configuration for pools of 3, 5, and 10 samples. Assessment of pooling using the TaqPath COVID-19 Combo Kit multiplex assay ( ORF1ab , N , and S gene targets) involved a limit of detection (LOD) study, matrix effect study, and clinical comparison of neat to pooled sample. An LOD of 135.02 ( ORF1ab ; CI.95: 117.21-155.52), 373.92 ( N ; CI.95: 257.05-437.64), and 1001.32 ( S ; CI.95: 896.62-1118.33) gce per milliliter was resolved. Seventy-two randomly selected samples demonstrated slight suppression due to negative sample matrix. Resulting mean cycle threshold (CT) shifts were 2.09 ( ORF1ab ), 1.76 ( N ), and 2.31 ( S ) for the 3-pool, 2.83 ( ORF1ab ), 2.45 ( N ), and 3.24 ( S ) for the 5-pool, and 3.99 ( ORF1ab ), 3.46 ( N ), and 4.07 ( S ) for the 10-pool. Despite quantitative sensitivity loss trend, the qualitative result was unaffected in each pool. According to the range of disease prevalence observed at the testing site (0.03-7.32%), a pool of five samples was deemed an optimal and cost-effective option for monitoring the Northeastern University community.

Population testing for severe acute respiratory syndrome 2 (SAR-CoV-2) is necessary owing to the possibility of viral transmission from asymptomatic cases, yet scarcity of reagents and equipment has added to the cost prohibitive implementation of screening campaigns at institutions of higher education. The high analytical sensitivities of leading nucleic acid amplification diagnostic methods allow for group testing to increase testing capacity. A feasibility study was performed using an optimized testing configuration model for pooling three, five, and ten samples. Following the standard RNA extraction and purification workflow for quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) method using Thermo Fisher TaqPath COVID-19 multiplex primers and probes for the ORF1ab, N, and S genes, matrix and dilution effects were assessed using pooled negative samples as the diluent. Probit analysis produced a limit of detection of 16075 (ORF1ab), 1308 (N), and 1180182 (S) genomic copy equivalents per milliliter. Trials comparing neat to 1:5 dilution for 34 weak-to-strongly positive samples demonstrated average threshold cycle (CT) shifts of 2.31+/-1.16 (ORF1ab), 2.23+/-1.12 (N), and 2.79+/-1.40 (S). Notwithstanding observed S gene dropouts, the false negative rate was unaffected. As the ratio of asymptomatic positive to symptomatic positive SARS-CoV-2 infected individuals was approximately 4:1 and the average prevalence was 0.16% since we started testing in August 2020, pooled testing was identified as a viable, cost-effective option for monitoring the Northeastern University community.