【病毒外文文獻(xiàn)】2018 Replicative virus shedding in the respiratory tract of patients with Middle East respiratory syndrome coronavirus i

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Accepted Manuscript Title Replicative virus shedding in the respiratory tract of the patients with Middle East respiratory syndrome coronavirus infection Authors Wan Beom Park Leo LM Poon Su Jin Choi Pyoeng Gyun Choe Kyoung Ho Song Ji Hwan Bang Eu Suk Kim Hong Bin Kim Sang Won Park Nam Joong Kim Malik Peiris Myoung don Oh PII S1201 9712 18 34411 4 DOI https doi org 10 1016 j ijid 2018 05 003 Reference IJID 3233 To appear in International Journal of Infectious Diseases Received date 21 3 2018 Revised date 3 5 2018 Accepted date 4 5 2018 Please cite this article as Park Wan Beom Poon Leo LM Choi Su Jin Choe Pyoeng Gyun Song Kyoung Ho Bang Ji Hwan Kim Eu Suk Kim Hong Bin Park Sang Won Kim Nam Joong Peiris Malik Oh Myoung don Replicative virus shedding in the respiratory tract of the patients with Middle East respiratory syndrome coronavirus infection International Journal of Infectious Diseases https doi org 10 1016 j ijid 2018 05 003 This is a PDF file of an unedited manuscript that has been accepted for publication As a service to our customers we are providing this early version of the manuscript The manuscript will undergo copyediting typesetting and review of the resulting proof before it is published in its final form Please note that during the production process errors may be discovered which could affect the content and all legal disclaimers that apply to the journal pertain Park et al 1 13 MS IJID D 18 00230 Replicative virus shedding in the respiratory tract of the patients with Middle East respiratory syndrome coronavirus infection Short communications Running Head Detection of MERS CoV subgenomic mRNA Wan Beom Park M D Ph D 1 2 Leo LM Poon 3 Su Jin Choi M Sc 2 Pyoeng Gyun Choe M D 1 Kyoung Ho Song M D Ph D 1 Ji Hwan Bang M D Ph D 1 Eu Suk Kim M D Ph D 1 Hong Bin Kim M D Ph D 1 Sang Won Park M D Ph D 1 Nam Joong Kim M D Ph D 1 2 Malik Peiris 3 and Myoung don Oh M D Ph D 1 2 1 Department of Internal Medicine Seoul National University College of Medicine Seoul South Korea 2 Laboratory of Infection RNA Messenger Respiratory System 1 Introduction Middle East respiratory syndrome coronavirus MERS CoV genomic RNA can persist for more than 1 month in respiratory specimens 1 However MERS CoV RNA detection may overestimate the duration of shedding of replicative virus Coronaviruses CoVs have a unique mechanism of discontinuous transcription with the synthesis of subgenomic mRNAs 2 The MERS CoV has at least seven distinct subgenomic mRNA species and detection of these indicates the virus is replicative 3 The objectives of this study was to examine the duration for detecting MERS CoV subgenomic mRNA vs genomic RNA in different respiratory specimens 2 Methods We collected respiratory samples from 17 patients admitted to three Seoul National University SNU affiliated hospitals during the 2015 MERS outbreak in Korea The patients were categorized into severe A I or mild J Q groups depending on their oxygen supplementation requirements The severe group required oxygen supplementation to maintain ACCEPTED MANUSCRIPT Park et al 4 13 arterial saturation above 90 percent Patients A E received ventilator therapy while patients F I did not 4 The institutional review board at SNU Hospital provided study approval and waived the requirement for written consent Oropharyngeal and nasopharyngeal swabs were collected by using UTMTM kit containing viral transport media Copan Diagnostics Inc Murrieta CA The RNA was extracted from respiratory samples using the QIAamp viral RNA mini kit QIAGEN Valencia CA To detect the MERS CoV genomic RNA multiplex quantitative real time reverse transcription rRT PCR was performed using the PowerChekTM MERS upE 29 307 8 2 Sawicki SG Sawicki DL Siddell SG A contemporary view of coronavirus transcription J Virol 2007 81 20 9 3 Woo PC Lau SK Fan RY Lau CC Wong EY Joseph S et al Isolation and characterization of dromedary camel coronavirus uae hku23 from dromedaries of the Middle East Minimal serological cross reactivity between MERS coronavirus and dromedary camel coronavirus uae hku23 Int J Mol Sci 2016 17 4 Park WB Perera RA Choe PG Lau EH Choi SJ Chun JY et al Kinetics of serologic responses to mers coronavirus infection in humans south korea Emerg Infect Dis 2015 21 2186 9 5 Oh MD Park WB Choe PG Choi SJ Kim JI Chae J et al Viral load kinetics of MERS coronavirus infection N Engl J Med 2016 375 1303 5 6 Corman VM Albarrak AM Omrani AS Albarrak MM Farah ME Almasri M et al Viral shedding and antibody response in 37 patients with Middle East respiratory syndrome coronavirus infection Clin Infect Dis 2016 62 477 83 7 Min CK Cheon S Ha NY Sohn KM Kim Y Aigerim A et al Comparative and kinetic analysis of viral shedding and immunological responses in mers patients representing a broad spectrum of disease severity Sci Rep 2016 6 25359 8 WHO Management of asymptomatic persons who are rt PCR positive for Middle East respiratory syndrome coronavirus MERS CoV Interim guidance Updated 3 jan 2018 Available at http apps who int iris bitstream 10665 180973 1 WHO MERS IPC 15 2 eng pdf u ACCEPTED MANUSCRIPT Park et al 8 13 a 1 ua 1 accessed March 21 2018 Figure Legends Fig 1 Detection of MERS CoV subgenomic mRNA in sputum or transtracheal aspirates A throat swabs B and nasopharyngeal swabs C A pink dot denotes a positive RT PCR result for MERS CoV subgenomic mRNA while a gray dot indicates a negative result The blue lines indicate the duration from the symptom onset to the last positive result for MERS CoV genomic RNA Patients A I were in the severe group and patients J Q were in the mild group Among the severe group subjects patients A E received ventilator therapy while patients F I did not The 5UTR S and 5UTR N denote the subgenomic mRNA of spike and nucleocapsid proteins respectively Fig 2 MERS CoV genomic RNA upE titers in sputum or transtracheal aspirates with vs without subgenomic mRNA detection The solid lines indicate means and standard errors of the mean The dashed line indicates the detection limit ACCEPTED MANUSCRIPT Park et al 9 13 Fig 1A ACCEPTED MANUSCRIPT Park et al 10 13 Fig 1B ACCEPTED MANUSCRIPT Park et al 11 13 Fig 1C ACCEPTED MANUSCRIPT Park et al 12 13 U n d e t e c t e d D e t e c t e d 3 4 5 6 7 8 9 10 p 0 0007 S u b g e n o m i c m RN A V i r a l RN A L o a d l o g 10 c o p i e s m l Fig 2 ACCEPTED MANUSCRIPT Park et al 13 13 Table 1 PCR primer pairs used in this study Forwa rd sequence 5 3 Reverse sequence 5 3 5 UT R S 1For GATTTAAGTGAATAGCT TGGCTA S21751 Rev TGAGAATAGTTAGCTACA AACAACT 5 UT R N 1For GATTTAAGTGAATAGCT TGGCTA N29121 Rev TCCTGATCTTGAACCTTGT GAACT ACCEPTED MANUSCRIPT