China CDC Weekly

Notes from the Field

First Human Infection with Avian H9N2 — Guangdong Province, China, 2020

Qianfang Guo1; Lirong Zou1; Jianxiang Yu1; Yingchao Song1; Lijun Liang1; Xue Zhuang1; Tie Song1; Jie Wu1,2,#

A case of H9N2 was detected in a 07/2018(H9N2) at 99.06%. Comparison with the NA human in Zhuhai City, Guangdong Province through genes of 36 H9N2 virus isolates from human infection routine influenza surveillance on March 30, 2020. The cases, sequence similarity was highest at 98.4% with patient was a 3-year-old child who developed fever and A/Fujian-Sanyuan/2881/2019(H9N2). cough on March 22. A pharyngeal swab was collected Phylogenetic analysis of HA and NA was carried on March 23, and the novel coronavirus nucleic acid with reference strains including the World Health test was negative. On March 26, the pharyngeal swab Organization (WHO) recommended pre-pandemic was sent to Zhuhai CDC for influenza virus tests and vaccine strains (1), similar H9N2 strain sequences was found to be H9 influenza A positive, and on from GenBank, and all human avian influenza virus March 30, Guangdong Provincial CDC reexamined sequences from GISAID. HA phylogenetic tree the pharyngeal swabs as positive for influenza A analysis showed the virus belong to the Y280/G9 clade (H9N2). On March 28, the patient was hospitalized and shared the same branch with A/duck/China/ for isolation treatment, and oseltamivir and other D4/2018(H9N2) from ducks and A/Hunan/34179/ antiviral drugs were given in the hospital. A pharyngeal 2018(H9N2) from humans (Figure 1). NA swab was collected again for reexamination on March Phylogenetic tree analysis showed the virus belongs to 28, and the H9 test was negative. Epidemiological the Y280/G9 clade and was clustered with A/chicken/ investigation found that the patient had been exposed Shanghai/07/2018(H9N2), A/chicken/China/1103/ to chickens and ducks raised by her grandparents 2019(H9N2), and A/chicken/China/71/2019(H9N2) before onset of illness. Samples taken from 5 close (2) (Figure 2). BLAST analysis of the internal gene contacts and the environment of her grandparents’ segments (MP, NP, NS, PA, PB1, PB2) of the isolated home all tested negative for the H9 nucleic acid. strain found that these 6 segments all had the highest The pharyngeal swab samples collected on March 23 similarity with avian-derived A (H9N2) with were inoculated into an 11-day-old specific pathogen molecular markers of avian-like viruses, suggesting that free (SPF) chicken embryo in Guangdong Provincial the risk of human infection was not higher than that of CDC. On April 7, the chicken embryo’s allantoic fluid previous H9N2 avian influenza viruses. was collected and identified as A(H9N2) positive, with H9N2 is the most common subtype of avian the strain named A/Guangdong/Zhuhai/20SF8034/ influenza in poultry, and human infection is relatively 2020(H9N2). The isolated strain was sequenced and rare (3–4). A total of 11 cases of H9N2 have been all 8 virus sequences of A(H9N2) were obtained. The 8 reported since 2010 in Guangdong Province, all of gene sequences have been uploaded to GISAID which were mild cases and have recovered. The case of (GISAID accession numbers: EPI1738873- H9N2 detected this time is an occasional case of EPI1738880). Basic local alignment search tool human infection with H9N2 avian influenza virus. We (BLAST) in GenBank showed that the hemagglutinin should continue to strengthen the risk monitoring and (HA) gene of this virus was most similar with early warning of avian influenza on the basis of the A/duck/China/d4/2018(H9N2) that was isolated from existing surveillance of avian influenza. Guangdong Province, China, with a sequence Fundings: This study was supported by the Key similarity of 99.06%. When compared with the HA Research and Development Program of Guangdong genes of 36 H9N2 virus isolates from human infection Province (2019B111103001), and Natural Science cases (searched from GISAID), the sequence similarity Foundation of Guangdong Province (2019A1515 with A/Hunan/34179/2018 was the highest (98.4%). 012121). BLAST in GenBank showed that the NA gene of this doi: 10.46234/ccdcw2020.147 virus was most similar with A/chicken/Shanghai/ # Corresponding author: Jie Wu, [email protected].

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A/Guangdong/Zhuhai/20SF8034/2020(H9N2) 96 A/duck/China/D4/2018(H9N2) 94 A/chicken/China/C7/2018(H9N2) 57 A/Hunan/34179/2018 A/chicken/Jiangxi/X1078/2018(H9N2) A/chicken/Hubei/H1081/2018(H9N2) A/chicken/Jiangxi/X2025/2017(H9N2) 77 A/chicken/Shanghai/S1182/2018(H9N2) 59 A/Duck/Jiangxi/X2083/2017(H9N2) A/chicken/Jiangxi/X2087/2017(H9N2) 94 52 A/Pigeon/Jiangxi/X2029/2017(H9N2) 94 A/chicken/Jiangxi/X2028/2017(H9N2) A/Fujian-Sanyuan/2881/2019 A/chicken/Huibei/H2244/2016(H9N2) 100 A/Duck/Huibei/H2247/2016(H9N2) A/chicken/Huibei/H2034/2016(H9N2) Y280/G9 86 A/chicken/Guangdong/G1301/2016(H9N2) A/environment/Hunankaifu/002/2017(H9N2) 66 A/chicken/Hubei/01/2015(H9N2) 73 A/Hunan/42088/2017 66 93 A/Hunan/37286/2017 58 A/Anhui-Lujiang/39/2018 A/Hunan/44558/2015 89 100 A/Hunan/44557/2015 27 A/Beijing/1/2017 99 A/Beijing/1/2016 A/Guangxi-Xiangshan/11522/2018 20 A/Guangdong/18SF064/2018 99 A/Hong Kong/623/2020 100 91 86 A/Guangdong/18SF003/2018 A/Hong Kong/308/2014 100 A/Zhongshan/201501/2015 96 A/Guangdong/MZ058/2016 A/Hong Kong/69955/2008 80 100 A/Hong Kong/3239/2008 72 A/Shaoguan/447/98 A/Shaoguan/408/98 93 A/Nanchang/D1/2000 66 A/Nanchang/D2/2000 99 A/Nanchang/CH3/2000 A/Nanchang/CH2/2000 A/Shantou/239/98 A/Guangzhou/333/99 A/HK/2108/2003 100 100 A/Guangdong/W1/2004 A/Korea/KBNP-0028/2000 100 A/Senegal/0243/2019 100 A/Oman/2747/2019 82 A/Bangladesh/0994/2011 100 A/Hong Kong/35820/2009 99 A/Hong Kong/33982/2009(2) 100 A/Hong Kong/1074/1999 84 A/Hong Kong/1073/99(2)

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FIGURE 1. Phylogenetic tree based on HA gene sequences of the isolated virus strain A/Guangdong/Zhuhai/20SF8034/ 2020(H9N2) and the reference strains using the maximum likelihood method. A red solid triangle marks the location of A/Guangdong/Zhuhai/20SF8034/2020(H9N2).

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A/Guangdong/Zhuhai/20SF8034/2020 43 A/chicken/Shanghai/07/2018(H9N2) 27 A/chicken/China/1103/2019(H9N2) 68 A/chicken/Shanghai/06/2018(H9N2) 91 A/chicken/China/71/2019(H9N2) 72 A/environment-air/Kunshan/NIOSH-BL25/2018(H9N2) 72 A/environment-air/Kunshan/NIOSH-BL20/2018(H9N2) A/chicken/Fujian/SD037/2017(H9N2) 86 A/Fujian-Sanyuan/2881/2019 60 24 A/Hong Kong/623/2020 69 A/Hong Kong/VB20030991/2020 97 A/Hunan/34179/2018 32 A/chicken/Japan/AQ-HE28-28/2016(H9N2) A/chicken/Shandong/k147/2017(H9N2) 71 A/chicken/Vietnam/HU9-447/2018(H9N2) 53 A/Guangdong/18SF003/2018 99 A/Guangxi-Xiangshan/11522/2018 Y280/G9 72 A/Guangdong/18SF064/2018 22 A/Anhui-Lujiang/39/2018 99 A/Hunan/42088/2017 5557 A/chicken/Anhui/AH120/2015(H9N2) 84 A/chicken/Zhejiang/HZ03/2014(H9N2) 23 A/chicken/Jiangxi/15492/2014(mixed) 82 A/Guangdong/MZ058/2016 81 A/chicken/JinShui/JS1002/2018(H9N2) 23 A/Hunan/37286/2017 94 A/chicken/Hubei/01/2015(H9N2) 23 A/Beijing/1/2016 A/Beijing/1/2017 80 A/Zhongshan/201501/2015 99 A/Hunan/44557/2015 99 A/Hunan/44558/2015 56 A/Hong Kong/308/2014 66 A/Shaoguan/408/98 A/Shaoguan/447/98 A/Nanchang/CH2/2000 100 29 A/Nanchang/CH3/2000 96 A/Nanchang/D1/2000 A/Nanchang/D2/2000 84 A/Bangladesh/0994/2011 46 A/Pakistan/486/2015 76 A/Oman/2747/2019 A/Hong Kong/1073/99 46 96 A/Hong Kong/1074/1999 A/Korea/KBNP-0028/2000 A/Shantou/239/98 A/Guangzhou/333/99 59 98 A/Hong Kong/3239/2008 61 A/Hong Kong/69955/2008 93 A/Guangdong/W1/2004 25 A/HK/2108/2003 93 A/Hong Kong/35820/2009 99 A/Hong Kong/33982/2009 83 A/Hong Kong/33982/2009(2) A/Senegal/0243/2019

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FIGURE 2. Phylogenetic tree based on neuraminidase (NA) gene sequences of the isolated virus strain A/Guangdong/Zhuhai/ 20SF8034/2020 (H9N2) and the reference strains using the maximum likelihood method. A red solid triangle marked the location of A/Guangdong/Zhuhai/20SF8034/2020(H9N2).

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2/en/.[2020-05-21]. 1 Guangdong Provincial Center of Disease Control and Prevention, 2. Wu YH, Lin JS, Yang SH, Xie Y, Wang M, Chen XQ, et al. The Guangzhou, Guangdong, China; 2 Southern Medical University, molecular characteristics of avian influenza viruses (H9N2) derived from School of Public Health, Guangzhou, Guangdong, China. air samples in live poultry markets. Infect Genet Evol 2018;60:191 − 6. http://dx.doi.org/10.1016/j.meegid.2018.01.009.

Submitted: June 24, 2020; Accepted: July 02, 2020 3. Li C, Wang SG, Bing GX, Carter RA, Wang ZJ, Wang JL, et al. Genetic evolution of influenza H9N2 viruses isolated from various hosts in China from 1994 to 2013. Emerg Microbes Infect 2017;6(1):e106. REFERENCES http://dx.doi.org/10.1038/emi.2017.94.

4. Xia J, Cui JQ, He X, Liu YY, Yao KC, Cao SJ, et al. Genetic and antigenic evolution of H9N2 subtype avian influenza virus in domestic

1. World Health Organization. Candidate vaccine viruses for A(H9N2). chickens in southwestern China, 2013-2016. PLoS One 2017;12(2): https://www.who.int/influenza/vaccines/virus/candidates_reagents/a_h9n e0171564. http://dx.doi.org/10.1371/journal.pone.0171564.

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