Diagnostic performance of RT-PCR-based sample pooling strategy for the detection of SARS-CoV-2

Miguel Hueda-Zavaleta, Cesar Copaja-Corzo, Vicente A. Benites-Zapata, Pedro Cardenas-Rueda, Jorge L. Maguiña, Alfonso J. Rodríguez-Morales

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Background: The rapid spread of SARS-CoV-2 has created a shortage of supplies of reagents for its detection throughout the world, especially in Latin America. The pooling of samples consists of combining individual patient samples in a block and analyzing the group as a particular sample. This strategy has been shown to reduce the burden of laboratory material and logistical resources by up to 80%. Therefore, we aimed to evaluate the diagnostic performance of the pool of samples analyzed by RT-PCR to detect SARS-CoV-2. Methods: A cross-sectional study of diagnostic tests was carried out. We individually evaluated 420 samples, and 42 clusters were formed, each one with ten samples. These clusters could contain 0, 1 or 2 positive samples to simulate a positivity of 0, 10 and 20%, respectively. RT-PCR analyzed the groups for the detection of SARS-CoV-2. The area under the ROC curve (AUC), the Youden index, the global and subgroup sensitivity and specificity were calculated according to their Ct values that were classified as high (H: ≤ 25), moderate (M: 26–30) and low (L: 31–35) concentration of viral RNA. Results: From a total of 42 pools, 41 (97.6%) obtained the same result as the samples they contained (positive or negative). The AUC for pooling, Youden index, sensitivity, and specificity were 0.98 (95% CI, 0.95–1); 0.97 (95% CI, 0.90–1.03); 96.67% (95% CI; 88.58–100%) and 100% (95% CI; 95.83–100%) respectively. In the stratified analysis of the pools containing samples with Ct ≤ 25, the sensitivity was 100% (95% CI; 90–100%), while with the pools containing samples with Ct ≥ 31, the sensitivity was 80% (95% CI, 34.94–100%). Finally, a higher median was observed in the Ct of the clusters, with respect to the individual samples (p < 0.001). Conclusions: The strategy of pooling nasopharyngeal swab samples for analysis by SARS-CoV-2 RT-PCR showed high diagnostic performance.

Original languageEnglish
Article number11
JournalAnnals of Clinical Microbiology and Antimicrobials
Issue number1
StatePublished - Dec 2022


  • COVID-19
  • COVID-19 diagnosis
  • Genetic techniques
  • Molecular diagnostic techniques
  • PCR
  • Peru (MeSH)
  • Pool testing
  • RT-PCR
  • SARS-CoV-2
  • Sensitivity and specificity


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