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Isolation and genetic characterization of SARS-CoV-2 from Indian patients in a single family without H/O travel abroad
Virus Genes ( IF 1.9 ) Pub Date : 2021-03-08 , DOI: 10.1007/s11262-021-01826-z
Shubham Shrivastava 1 , Harshad P Patil 1 , Suhas T Mhaske 1 , Sonali Palkar 2 , Sanjay Lalwani 2 , Akhilesh Chandra Mishra 1 , Vidya A Arankalle 1
Affiliation  

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 × 106 and 4 × 106 Pfu/ml by plaque assay and 107.5 and 107 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.



中文翻译:

从没有 H/O 出国旅行的单个家庭的印度患者中分离和遗传表征 SARS-CoV-2

鉴于迅速发展的 COVID-19 大流行,我们的目标是从印度患者中分离和表征 SARS-CoV-2。SARS-CoV-2 是从一个没有任何 (H/O) 出国旅行史的家庭的两名成员采集的鼻咽拭子中分离出来的。两种病毒分离物(8003 和 8004)在接种后第 3 天显示 CPE,通过免疫荧光测定显示病毒抗原,并产生明显、清晰和均匀的斑块。通过噬斑测定,传染性病毒滴度为 5 × 10 6和 4 × 10 6 Pfu/ml,10 7.5和 10 7分别通过基于 CPE 的 TCID50/ml。系统发育分析将我们的分离株与意大利菌株分组。与武汉株相比,分别在Orf1ab的nsp3(A1812D)、核酸外切酶(P1821S)和刺突蛋白(Q677H)区域鉴定出3个独特突变。这两种病毒都与意大利的 SARS-CoV-2 毒株分组,这表明可能的来源是从意大利进口的病毒。这些完全表征的病毒分离物将有助于开发用于评估疫苗/抗病毒药物的中和/病毒学分析。

更新日期:2021-03-08
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