SARS-CoV-2 isolation and propagation from Turkish COVID-19 patients

The novel coronavirus pneumonia, which was named later as coronavirus disease 2019 (COVID-19), is caused by the severe acute respiratory syndrome coronavirus 2, namely SARS-CoV-2. It is a positive-strand RNA virus that is the seventh coronavirus known to infect humans. The COVID-19 outbreak presents enormous challenges for global health behind the pandemic outbreak. The first diagnosed patient in Turkey has been reported by the Republic of Turkey Ministry of Health on March 11, 2020. In May, over 150,000 cases in Turkey, and 5.5 million cases around the world have been declared. Due to the urgent need for a vaccine and antiviral drug, isolation of the virus is crucial. Here, we report 1 of the first isolation and characterization studies of SARS-CoV-2 from nasopharyngeal and oropharyngeal specimens of diagnosed patients in Turkey. This study provides an isolation and replication methodology,and cell culture tropism of the virus that will be available to the research communities.

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Andersen KG, Rambaut A, Lipkin WI, Holmes EC, Garry RF (2020). The proximal origin of SARS-CoV-2. Nature Medicine 26: 450- 52. doi: 10.1038/s41591-020-0820-9
Baer A, Kylene KH (2014). Viral concentration determination through plaque assays: using traditional and novel overlay systems. Journal of Visualized Experiments 93: 1-10. doi: 10.3791/52065
Cui J, Fang L, Zheng LS (2019). Origin and evolution of pathogenic coronaviruses. Nature Reviews Microbiology 17: 181-192. doi: 10.1038/s41579-018-0118-9
Guallart N (2015). An overview of virus qunatification techniques. Axiomathes 25: 61-77. doi: 10.1007/s10516-014-9260-9
Harcourt J, Tamin A, Lu X, Kamili S, Kumar S (2020). Isolation and characterization of SARS-CoV-2 from the first US COVID-19 patient. bioRxiv doi: https://doi.org/10.1101/2020.03.02.972935
He L, Ding Y, Zhang Q, Che X, He Y et al. (2006). Expression of elevated levels of pro-inflammatory cytokines in SARS-CoVinfected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS288–97. The Journal of Pathology 210 (3): 288-297. doi: 10.1002/path
Holshue ML, DeBoltC, Lindquist S, Lofy KH, Wiesman J et al. (2020). First case of 2019 novel coronavirus in the United States. The New England Journal of Medicine 382 (10): 929- 936. doi: 10.1056/NEJMoa2001191
Honda-Okubo Y, Barnard D, Ong CH, Peng BH, Kent Tseng CT et al. (2015). Severe acute respiratory syndrome-associated coronavirus vaccines formulated with delta inulin adjuvants provide enhanced protection while ameliorating lung eosinophilic immunopathology. Journal of Virology 89 (6): 2995-3007. doi: 10.1128/jvi.02980-14
Hui DS, Azhar EI, Madani TA, Ntoumi F, Kock R (2020). The continuing 2019-NCoV epidemic threat of novel cCoronaviruses to global health — the latest 2019 novel coronavirus outbreak in Wuhan, China. International Journal of Infectious Diseases 91: 264-266. doi: 10.1016/j. ijid.2020.01.009
Li CK, Wu H, Yan H, Ma S, Wang L et al. (2008). T cell responses to whole SARS coronavirus in humans.The Journal of Immunology 181 (8): 5490-5500. doi: 10.4049/jimmunol.181.8.5490
Li Q, Guan X, Wu P, Wang X, Zhou L et al. (2020). Early transmission dynamics in Wuhan, China, of novel coronavirus-infected pneumonia. The New England Journal of Medicine 382 (13): 1199-1207. doi: 10.1056/NEJMoa2001316
Libraty DH, O’Neil KM, Baker LM, Acosta LP, Olveda RM (2007). Human CD4+ memory T-lymphocyte responses to SARS coronavirus infection.Virology 368 (2): 317-321. doi: 10.1016/j. virol.2007.07.015
Liu W, Li H (2020). COVID-19: Attacks the 1-beta chain of hemoglobin and captures the porphyrin to inhibit human meme metabolism. ChemRxiv. doi: 10.26434/chemrxiv
Masters PS, Perlman S (2013). Coronaviridae. In: Knipe DM, Howley PM, Cohen JI, Griffen DE, Lamb RA et al. (editors). Fields Virology, Vol. 1. 6th ed. Philadelphia, PA, USA: Lippincott Williams & Wilkins, pp. 825-858.
Matsumoto M, Mukai T, Furukawa S, Ohori H (2005). Inhibitory effects of epigallocatechin gallate on the propagation of bovine coronavirus in Madin-Darby bovine kidney cells. Animal Science Journal 76 (5): 507-512. doi: 10.1111/j.1740- 0929.2005.00297.x
Paraskevis D, Kostaki EG, Magiorkinis G, Panayiotakopoulos G, Sourvinos G et al. (2020). Full-genome evolutionary analysis of the novel corona virus (2019-NCoV) rejects the hypothesis of emergence as a result of a recent recombination event. Infection, Genetics and Evolution 79: 1004212. doi: 10.1016/j. meegid.2020.104212
Stohl W, Mayer L (1987). Inhibition of T cell-dependent human B cell proliferation and B cell differentiation by polyspecific monomeric IgG. Clinical and Experimental Immunology 70 (3): 649-57.
Weiss SR, Leibowitz JL (2011). Chapter 4 - Coronavirus pathogenesis. Advances in Virus Research 81: 85-164. doi: 10.1016/B978-0- 12-385885-6.00009-2
Wong G, Liu W, Liu Y, Zhou B, Bi Y et al. (2015). MERS, SARS, and Ebola: The Role of Super-Spreaders in Infectious Disease. Cell Host & Microbe 18 (4): 398-401. doi: 10.1016/j. chom.2015.09.013
Zheng J (2020). SARS-CoV-2: An emerging coronavirus that causes a global threat. International Journal of Biological Sciences 16 (10): 1678-1685. /doi: 10.7150/ijbs.45053
Zhou P, Yang XL, Wang XG, Hu B, Zhang L et al. (2020). A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature 579 (7798): 270-273. /doi: 10.1038/s41586-020- 2012-7