BRIEF REPORT a powerful tool for the discovery of patho- Next-generatic sequencing and bioin can our one ance an nd e China.M nt dat understanding of disease occurrence and trans. show 2019-nCov detection in 830 persons in mission,accelerating the identification of patho-China. Although our study does not fulfill Koch's postul 01 nce im novel beracoronirus that is likely to have been ditional evidence to confirm the etiologic sig the cause of severe pneumonia in three patients nificance of 2019-nCoV in the Wuhan outbreak clude identification of a 2019-nCov antigen in is g human airway epith ng tissue of patients man respiratory pathogens.Our study showed a patient at two time points to demonstrate se- that initial propagation of human respiratory se roconversion,and animal (monkey)experiments ce to provide evidence of pat ogenicity.Of critica of culture characterize tra ssion modes. tion in terval,and clinical spectrum resulting from infec tion and detection of new human coronavirus tion to inform and refine strategies that can pre that can possibly elude identification by tradi- vent,control,and stop the spread of 2019-nCov of accurate and rapid methods to identify unknown re l Maio n of gens is still needed.On the basis of analysis of three complete genomes obtained in this study. w th of this the as ue He,Dr.L ce Kod nd E 10 Yu Li nCo genome to detect viral RNA in cinical ne fo ration and s specimens.The primer sets and standard oper- o GE. .G 3.0 shost inter rome.N E Med20034-1967. uceJ.Du Let al.A ne Wilkins..2013.82 acnmrccaNEngljMcd20o8 ie and 15318398-40 g NS,Zhe guM,tal.pi Ith.N 91231 icipal Heat Co Viral and al e 77 N ENGLJ MED NM.ORC n engl j med nejm.org 7 Brief Report a powerful tool for the discovery of patho￾gens.14,16 Next-generation sequencing and bioin￾formatics are changing the way we can respond to infectious disease outbreaks, improving our understanding of disease occurrence and trans￾mission, accelerating the identification of patho￾gens, and promoting data sharing. We describe in this report the use of molecular techniques and unbiased DNA sequencing to discover a novel betacoronavirus that is likely to have been the cause of severe pneumonia in three patients in Wuhan, China. Although establishing human airway epithe￾lial cell cultures is labor intensive, they appear to be a valuable research tool for analysis of hu￾man respiratory pathogens.13 Our study showed that initial propagation of human respiratory se￾cretions onto human airway epithelial cell cul￾tures, followed by transmission electron micros￾copy and whole genome sequencing of culture supernatant, was successfully used for visualiza￾tion and detection of new human coronavirus that can possibly elude identification by tradi￾tional approaches. Further development of accurate and rapid methods to identify unknown respiratory patho￾gens is still needed. On the basis of analysis of three complete genomes obtained in this study, we designed several specific and sensitive assays targeting ORF1ab, N, and E regions of the 2019- nCoV genome to detect viral RNA in clinical specimens. The primer sets and standard oper￾ating procedures have been shared with the World Health Organization and are intended for surveillance and detection of 2019-nCoV infec￾tion globally and in China. More recent data show 2019-nCoV detection in 830 persons in China.17 Although our study does not fulfill Koch’s postulates, our analyses provide evidence impli￾cating 2019-nCoV in the Wuhan outbreak. Ad￾ditional evidence to confirm the etiologic sig￾nificance of 2019-nCoV in the Wuhan outbreak include identification of a 2019-nCoV antigen in the lung tissue of patients by immunohisto￾chemical analysis, detection of IgM and IgG antiviral antibodies in the serum samples from a patient at two time points to demonstrate se￾roconversion, and animal (monkey) experiments to provide evidence of pathogenicity. Of critical importance are epidemiologic investigations to characterize transmission modes, reproduction in￾terval, and clinical spectrum resulting from infec￾tion to inform and refine strategies that can pre￾vent, control, and stop the spread of 2019-nCoV. This work was supported by grants from the National Key Research and Development Program of China (2016YFD0500301) and the National Major Project for Control and Prevention of Infectious Disease in China (2018ZX10101002). Disclosure forms provided by the authors are available with the full text of this article at NEJM.org. We thank Dr. Zhongjie Li, Dr. Guangxue He, Dr. Lance Rode￾wald, Yu Li, Fei Ye, Li Zhao, Weimin Zhou, Jun Liu, Yao Meng, Huijuan Wang, and many staff members at the China CDC for their contributions and assistance in this preparation and sub￾mission of an earlier version of the manuscript. References 1. Gao GF. From “A”IV to “Z”IKV: attacks from emerging and re-emerging patho￾gens. Cell 2018;172:1157-9. 2. Weiss SR, Leibowitz JL. Coronavirus pathogenesis. Adv Virus Res 2011;81:85- 164. 3. Masters PS, Perlman S. Coronaviridae. In: Knipe DM, Howley PM, eds. Fields vi￾rology. 6th ed. Lippincott Williams & Wilkins, 2013:825-58. 4. Su S, Wong G, Shi W, et al. Epidemiol￾ogy, genetic recombination, and patho￾genesis of coronaviruses. Trends Micro￾biol 2016;24:490-502. 5. Cui J, Li F, Shi ZL. Origin and evolu￾tion of pathogenic coronaviruses. Nat Rev Microbiol 2019;17:181-92. 6. Zhong NS, Zheng BJ, Li YM, et al. 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