Ready for new waves: optimizing SARS-CoV-2 variants monitoring in pooled samples with droplet digital PCR

为新一轮疫情做好准备:利用液滴数字 PCR 优化混合样本中 SARS-CoV-2 变异株的监测

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作者:Antonella Pacini #, Franco Paredes #, Sofia Heckel, Guadalupe Ibarra, Maria Victoria Petreli, Marilina Perez, Yanina Agnella, Laura Piskulic, Maria Belen Allasia, Luis Caprile, Alejandro Colaneri, Juliana Sesma

Discussion

The employment of two separate TaqMan probes for both Delta and Omicron establishes dual validation routes, emphasizing the method's robustness. Although we used known samples to model realistic emergence scenarios of the Delta and Omicron variants, our main objective is to demonstrate the versatility of this strategy to identify future variant appearances. The utilization of two divergent variants and distinct probes for each confirms the method's independence from specific variants and probes. This flexibility ensures it can be tailored to recognize any subsequent variant emergence, given the availability of its sequence and a specific probe. Consequently, our approach stands as a robust tool for tracking and managing any new variant outbreak, reinforcing our global readiness against possible future SARS-CoV-2 waves.

Methods

Using 1200 retrospective SARS-CoV-2 positive samples, either negative or positive for Delta or Omicron, we assessed the sensitivity and specificity of our detection strategy employing commercial TaqMan variant probes in a 1:9 ratio of variant-positive to variant-negative samples.

Results

The study achieved 100% sensitivity and 99% specificity in 10-sample pools, with an Area Under the Curve (AUC) exceeding 0.998 in ROC curves, using distinct commercial TaqMan variant probes.

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