Nasopharyngeal samples (NPS) taken from people at any of the five hospitals of Central Greece were submitted for detection of SARs-CoV-2 and were processed immediately. Viral RNA was extracted from 400 μL from each NPS by using the commercial kit MagDEA®®Dx SV using a magLEAD®® 12gC instrument (Precision System Science Co, Matsudo city, Chiba, Japan). Detection of SARS-CoV-2 was performed by rRT-PCR, by using a commercial kit that targeted the E (common for other SARS-related coronaviruses) and N (specific for SARS-CoV-2) genes (Direct SARS-CoV-2 Real-Time PCR kit, Vircell, Granada, Spain), with a threshold limit of detection of 3.5 copies per reaction for both genes. The RNase P gene region was used as an endogenous internal control for the analysis of biological samples (Direct SARS-CoV-2 Real-Time PCR kit, Vircell, Granada, Spain).

A sample was considered to be SARS-CoV-2 positive, when Ct values for both the E and N genes were found to be <40, according to the recommendations of the manufacturer. In addition, samples in which Ct values for the RNAse P gene were found to be ≥40, were rejected.

After testing, all the NPS and RNAs were kept at −20 °C and −80 °C, respectively.

A copy of the final result of the test for each sampled person was sent to the referring clinicians, who were responsible for informing the people. The result of the test was also added into the Greek national platform e-Government Center for Social Security (IDIKA), as required by the national policy on the measures against COVID-19.

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