Department of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed To Be University), Katraj-Dhankawadi, Pune, 411043, India.
Department of Pediatrics, Bharati Vidyapeeth (Deemed To Be University) Medical College & Hospital, Katraj-Dhankawadi, Pune, 411043, India.
Virus Genes. 2021 Jun;57(3):245-249. doi: 10.1007/s11262-021-01826-z. Epub 2021 Mar 8.
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 and 4 × 10 Pfu/ml by plaque assay and 10 and 10 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.
鉴于 COVID-19 大流行迅速发展，我们的目的是从印度患者中分离和鉴定 SARS-CoV-2。从没有出国旅行史（H/O）的两个家庭成员的鼻咽拭子中分离出了 SARS-CoV-2。两种病毒分离株（8003 和 8004）在接种后第 3 天均出现 CPE，免疫荧光法检测到病毒抗原，并产生了明显、清晰和均匀的蚀斑。通过蚀斑测定法和基于 CPE 的 TCID50/ml 分别测定的感染性病毒滴度分别为 5×10 和 4×10 Pfu/ml。系统发育分析将我们的分离株与意大利株聚为一组。与武汉株相比，在 nsp3（A1812D）、ORF1ab 区的核酸外切酶（P1821S）和刺突蛋白（Q677H）区分别鉴定出 3 个独特的突变。两种病毒均与意大利 SARS-CoV-2 株聚为一组，提示可能的来源是从意大利输入的病毒。这些充分鉴定的病毒分离株将有助于开发中和/病毒学检测方法，用于评估疫苗/抗病毒药物。
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