RT-PCR:
All samples were subjected to RNA extraction with Qiagen Viral Nucleic
Acid Kit (QIAcub HT) according to the manufacturer’s instructions.
RT-PCR was used to detect the presence of SARS-CoV2 using the kits
(Molbiol, Germany) provided by WHO targeting the E region for screening
and RNA dependent RNA polymerase for confirmation. Invitrogen
Superscript III One-Step RT-PCR System with Platinum Taq DNA Polymerase
was used for PCR. For each reaction, 15 µl reaction mix, 1 µl RT enzyme,
0.5 µl primer, probe mix and 3.5 µl PCR grade water were added to 5 µl
RNA template. Cycling conditions for amplification of E and RdRP genes
were 50°C for 30 min, 95°C for 2 min, followed by 45 cycles of 95°C for
10 sec and 60°C for 30 sec. A cycle threshold value of <36 Ct
was defined as a positive test result.