Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing
Daniel Cruceriu,
Oana Baldasici,
Loredana Balacescu,
Stefana Gligor-Popa,
Mirela Flonta,
Milena A. Man,
Simona Visan,
Catalin Vlad,
Adrian P. Trifa,
Ovidiu Balacescu,
Patriciu Achimas-Cadariu
Affiliations
Daniel Cruceriu
Department of Genetics, Genomics and Experimental Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Oana Baldasici
Department of Genetics, Genomics and Experimental Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Loredana Balacescu
Department of Genetics, Genomics and Experimental Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Stefana Gligor-Popa
Department of Oncogenetics, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Mirela Flonta
Laboratory of Microbiology, Clinical Hospital of Infectious Diseases, 23, Iuliu Moldovan Street, 400348 Cluj-Napoca, Romania
Milena A. Man
Department of Medical Sciences—Pulmonology, “Iuliu Hatieganu” University of Medicine and Pharmacy, 8, Victor Babes Street, 400012 Cluj-Napoca, Romania
Simona Visan
Department of Genetics, Genomics and Experimental Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Catalin Vlad
Department of Surgery, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Adrian P. Trifa
Department of Oncogenetics, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Ovidiu Balacescu
Department of Genetics, Genomics and Experimental Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
Patriciu Achimas-Cadariu
Department of Surgery, The Oncology Institute “Prof. Dr. Ion Chiricuta”, 34-36, Republicii Street, 400015 Cluj-Napoca, Romania
The primary approach to controlling the spread of the pandemic SARS-CoV-2 is to diagnose and isolate the infected people quickly. Our paper aimed to investigate the efficiency and the reliability of a hierarchical pooling approach for large-scale PCR testing for SARS-CoV-2 diagnosis. To identify the best conditions for the pooling approach for SARS-CoV-2 diagnosis by RT-qPCR, we investigated four manual methods for both RNA extraction and PCR assessment targeting one or more of the RdRp, N, S, and ORF1a genes, by using two PCR devices and an automated flux for SARS-CoV-2 detection. We determined the most efficient and accurate diagnostic assay, taking into account multiple parameters. The optimal pool size calculation included the prevalence of SARS-CoV-2, the assay sensitivity of 95%, an assay specificity of 100%, and a range of pool sizes of 5 to 15 samples. Our investigation revealed that the most efficient and accurate procedure for detecting the SARS-CoV-2 has a detection limit of 2.5 copies/PCR reaction. This pooling approach proved to be efficient and accurate in detecting SARS-CoV-2 for all samples with individual quantification cycle (Cq) values lower than 35, accounting for more than 94% of all positive specimens. Our data could serve as a comprehensive practical guide for SARS-CoV-2 diagnostic centers planning to address such a pooling strategy.
