SARS-CoV-2 Pooled Testing Methodology for PCR Testing Applied in Private Laboratory in Armenia

Since the beginning of COVID-19 pandemics many countries were facing challenges with testing capacity recourse limitations. Throughout the waves of the pandemic countries were trying to address the existing constrains exploring solutions to increase the testing capacity with more cost-effective approaches. Pooled methodology was one of the methods which many have validated and used. It is evident that in case of pooled sample testing the sensitivity becomes lower, however the variation highly depends on the pool size as well as the incidence rate at the certain point. Armenia as well as many other countries has adopted regulations for mandatory COVID-19 PCR testing for all the travelers. Current study aimed to explore the efficiency of COVID-19 pooled PCR testing for nasopharyngeal swabs of individuals with no symptoms in a time period with good epidemiological state of the infection. Nasopharingeal swab samples from individuals were collected. The manual extraction of RNAs of samples was performed after pooling up to 5 samples. The pools with Cycle Threshold (CT) of < 37 were considered positive and were retested individually. In total 28,015 samples were grouped in 667 pools of which 57 were positive. The total number of positive samples was 65. The median difference (CT-pool–CT samples) was 2.4 (ranging from–3.0 to 8.9). The correlation of CT of pools and positive samples was positive. The correlation coefficient r = 0.84, P < 0.000, 95% CI range 0.7423 to 0.9243). The total economic saving when using pools compared to the individual testing was 72%. The minor difference between CT values of pools and samples can be explained by the dilution effect in the pool. However, the positive correlation between the values as well as the amount of cost saving demonstrate that pooling on nasopharyngeal samples for COVID-19 PCR testing can be a good method for efficient screening with significant resource saving. One of the most important advantages of the proposed method is the fact that samples are pooled prior extraction, which avoids the possibilities with misinterpretation of IC due to low yield of RNA in the extraction process.