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A Territory-Wide Study of Early COVID-19 Outbreak in Hong Kong Community: A

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A Territory-Wide Study of Early COVID-19 Outbreak in Hong Kong Community: A medRxiv preprint doi: https://doi.org/10.1101/2020.03.30.20045740; this version posted April 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. 1 A territory-wide study of early COVID-19 outbreak in Hong Kong community: A 2 clinical, epidemiological and phylogenomic investigation 3 4 Kenneth Siu-Sing LEUNG, Timothy Ting-Leung NG, Alan Ka-Lun WU, Miranda 5 Chong-Yee YAU, Hiu-Yin LAO, Ming-Pan CHOI, Kingsley King-Gee TAM, Lam- 6 Kwong LEE, Barry Kin-Chung WONG, Alex Yat-Man HO, Kam-Tong YIP, Kwok- 7 Cheung LUNG, Raymond Wai-To LIU, Eugene Yuk-Keung TSO, Wai-Shing LEUNG, 8 Man-Chun CHAN, Yuk-Yung NG, Kit-Man SIN, Kitty Sau-Chun FUNG, Sandy Ka- 9 Yee CHAU, Wing-Kin TO, Tak-Lun QUE, David Ho-Keung SHUM, Shea Ping YIP, 10 Wing Cheong YAM, Gilman Kit-Hang SIU1 11 12 The Hong Kong Polytechnic University, Hong Kong Special Administrative Region, China (T.NG, H-Y.LAO, 13 M-P.CHOI, L-K.LEE, D.SHUM, SP. YIP, G.SIU) 14 The University of Hong Kong, Hong Kong Special Administrative Region, China (K.LEUNG, K.TAM, W- 15 C.YAM) 16 Pamela Youde Nethersole Eastern Hospital, Hong Kong Special Administrative Region, China (A.WU, M.YAU, 17 K-C.LUNG) 18 Ruttonjee Hospital, Hong Kong Special Administrative Region, China (R.LIU) 19 United Christian Hospital, Hong Kong Special Administrative Region, China (B.WONG, E.TSO, K.FUNG, 20 S.CHAU) 21 Princess Margaret Hospital, Hong Kong Special Administrative Region, China (A.HO, W-S.LEUNG, M- 22 C.CHAN, W-K.TO) 23 Tuen Mun Hospital, Hong Kong Special Administrative Region, China (K-T.YIP, Y-Y.NG, K-M.SIN, T- 24 L.QUE) 25 26 1 Correspondence to: Dr. Gilman KH SIU, Department of Health Technology and Informatics, Hong 27 Kong Polytechnic University, Hong Kong ([email protected]) 28 29 30 Article summary lines: A combined epidemiological and phylogenetic analysis of early 31 COVID-19 outbreak in Hong Kong revealed that a SARS-CoV-2 variant with ORF3a G251V 32 mutation accounted for all locally acquired cases, and that asymptomatic carriers could be a 33 huge public health risk for COVID-19 control. 34 1 NOTE: This preprint reports new research that has not been certified by peer review and should not be used to guide clinical practice. medRxiv preprint doi: https://doi.org/10.1101/2020.03.30.20045740; this version posted April 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. 35 Running title: Early community outbreak of COVID-19 in Hong Kong 36 37 Keywords: Novel Coronavirus, COVID-19, SARS-CoV-2, community outbreak, whole 38 genome sequencing, clinical demographic, molecular phylogeny, molecular evolutionary 39 analysis 40 2 medRxiv preprint doi: https://doi.org/10.1101/2020.03.30.20045740; this version posted April 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. 41 Abstract (148 words) 42 43 Initial cases of COVID-19 reported in Hong Kong were mostly imported from China. 44 However, most cases reported in February 2020 were locally-acquired infections, indicating 45 local community transmissions. We extracted the demographic, clinical and epidemiological 46 data from 50 COVID-19 patients, who accounted for 53.8% of the cases in Hong Kong by 47 February 2020. Whole-genome sequencing of the SARS-CoV-2 were conducted to determine 48 the phylogenetic relatedness and transmission dynamics. Only three (6.0%) patients required 49 ICU admission. Phylogenetic analysis identified six transmission clusters. All locally- 50 acquired cases harboured a common mutation Orf3a G251V and were clustered in two 51 subclades in global phylogeny of SARS-CoV-2. The estimated time to the most recent 52 common ancestor of local COVID-2019 outbreak was December 24, 2019 with an 53 evolutionary rate of 3.04×10-3 substitutions per site per year. The reproduction number value 54 was 1.84. Social distancing and vigilant epidemiological control are crucial to the 55 containment of COVID-19 transmission. 56 57 3 medRxiv preprint doi: https://doi.org/10.1101/2020.03.30.20045740; this version posted April 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. 58 (3452 words) 59 Introduction 60 “Coronavirus disease 2019(COVID-19)” refers to a cluster of viral pneumonia cases initially 61 from Wuhan, Hubei Province, China since December 2019. The aetiology was unknown 62 during the early stage of the outbreak until Chinese scientists isolated a novel coronavirus, 63 SARS-CoV-2, on January 7, 2020 and performed genome sequencing(1). 64 Fever was the main symptom of COVID-19 with about one-third of the patients presenting 65 acute respiratory distress syndrome(ARDS). About 16% of the patients were in severe 66 condition on admission, and the estimated mortality rate was 1.4%(2). Sustained human-to- 67 human transmission was confirmed upon identification of cases clustering among families 68 and transmission from patients to healthcare workers(3, 4), which triggered China's urgent 69 public health actions and international concern. 70 As of February 28, 2020, 78,824 patients have been diagnosed to have COVID-19 in 71 Mainland China with 2,788 deaths. The disease also spread to 50 other countries (5). The 72 World Health Organization(WHO) declared COVID-19 pandemic in March 2020. The first 73 imported case in Hong Kong was on January 23, 2020, who was a Mainland China resident 74 travelled to Hong Kong from Wuhan via Shenzhen by High-Speed Rail. The first local case 75 with unknown source, i.e. patient who had no travel record during the 14-day incubation 76 period, was reported on February 4, 2020(6). 77 By February 28, 2020, there were 93 COVID-19 cases in Hong Kong with at least 70(75·3%) 78 cases being local cases and their close contacts(6, 7). Secondary and tertiary transmissions 79 were observed in some case clusters. As most index cases of these clusters have unknown 80 sources, hidden transmission chain was believed to have established in the community. 4 medRxiv preprint doi: https://doi.org/10.1101/2020.03.30.20045740; this version posted April 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. 81 Here, we report the demographic, clinical and epidemiological data of 50 hospitalised 82 patients, who accounted for 53.8% of COVID-19 cases in Hong Kong at the data cut-off 83 point (February 28, 2020), including three imported cases and six transmission clusters of 84 local infections. With the combined use of Nanopore sequencing and Illumina sequencing, 85 viral genomes of all these cases were characterised. Phylogenetic and molecular evolutionary 86 analyses were performed to determine the transmission linkage and the evolutionary rate of 87 COVID-19 cases in the community. 88 89 Methods 90 Cases 91 For this retrospective, multi-centre study, we intentionally enrolled the positive cases of 92 COVID-19 that were laboratory-confirmed at four public hospital clusters managed under the 93 Hospital Authority of Hong Kong, namely Hong Kong East Cluster, Kowloon East Cluster, 94 Kowloon West Cluster, and New Territories West Cluster, from January 26 to February 28, 95 2020. 96 Sputum specimens and throat swabs pooled with nasopharyngeal aspirates were collected 97 from patients who fulfilled the reporting or enhanced surveillance criteria on admission(8). 98 Laboratory-confirmed infection was defined when SARS-CoV-2 was detected by real-time 99 reverse transcription polymerase chain reactions(RT-PCRs), which amplified the envelope(E) 100 gene and RNA-dependent RNA polymerase(RdRp) gene(9). 101 Demographic, clinical, and microbiological data were obtained from patients’ medical 102 records. Epidemiological information was retrieved from the Centre for Health 103 Protection(CHP) of the Department of Health(6) and the website wars.vote4.hk - Coronavirus 104 in HK(7). The definitions of clinical symptoms and complications are based on the WHO 5 medRxiv preprint doi: https://doi.org/10.1101/2020.03.30.20045740; this version posted April 7, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. 105 guidance(10). We adopted the case numbering system of CHP, which was based on the date 106 of case confirmation. 107 This study was approved by the Institutional Review Boards of The Hong Kong Polytechnic 108 University(RSA20021) and the public hospitals involved(HKECREC-20200014;KCC/KEC- 109 20200070;KWC-20200040;NTWC-20200038). 110 111 Specimen preparation 112 The respiratory specimens were centrifuged at 16,000g for 2 minutes. Total nucleic acid was 113 extracted from supernatant using MagNA Pure 96 System(Roche Diagnostics, Germany) or 114 NucliSENS® easyMAG®(bioMérieux, The Netherlands) according to manufacturers’ 115 instructions.
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