Virus Research ( IF 2.5 ) Pub Date : 2020-09-24 , DOI: 10.1016/j.virusres.2020.198173 Byron Freire-Paspuel , Patricio Vega-Mariño , Alberto Velez , Marilyn Cruz , Miguel Angel Garcia-Bereguiain
Background
The CDC protocol for SARS-CoV2 RT-PCR diagnosis (2019-nCoV CDC kit) is considered a gold standard worldwide; based on three different FAM probes (N1 and N2 for viral detection; RP for RNA extraction quality control), three reactions per sample are needed for SARS-CoV-2 diagnosis.
Results
We herein describe a sample pooling protocol: pooling 3 RNA extractions into a single PCR reaction; we tested this protocol with 114 specimens grouped in 38 pools and found no significant differences for N1 and N2 Ct values between pool and single sample PCR reaction.
Conclusion
This pool of three protocol has a sensitivity of 100 % compared to the standard single sample protocol. For a typical 96-well plate, this pool assay allows 96 samples processing, speeding up diagnosis and reducing cost while maintaining clinical performance, particularly useful for SARS-CoV-2 diagnosis at developing countries.