Isolation and genetic characterization of SARS-CoV-2 from Indian patients in a single family without H/O travel abroad

Shrivastava, Shubham; Patil, Harshad P.; Mhaske, Suhas T.; Palkar, Sonali; Lalwani, Sanjay; Mishra, Akhilesh Chandra; Arankalle, Vidya A.

In view of the rapidly progressing COVID-19 pandemic, our aim was to isolate and characterize SARS-CoV-2 from Indian patients. SARS-CoV-2 was isolated from nasopharyngeal swabs collected from the two members of a family without any history of (H/O) travel abroad. Both the virus isolates (8003 and 8004) showed CPE on day 3 post-inoculation, viral antigens by immunofluorescence assay and produced distinct, clear and uniform plaques. Infectious virus titers were 5 × 10(6) and 4 × 10(6) Pfu/ml by plaque assay and 10(7.5) and 10(7) by CPE-based TCID50/ml, respectively. Phylogenetic analysis grouped our isolates with the Italian strains. On comparison with Wuhan strain, 3 unique mutations were identified in nsp3 (A1812D), exonuclease (P1821S) of Orf1ab and spike protein (Q677H) regions, respectively. Both the viruses grouped with Italian strains of SARS-CoV-2 suggesting possible source being the virus imported from Italy. These fully characterized virus isolates will be useful in developing neutralization/virological assays for the evaluation of vaccines/antivirals.