title: Improved detection of Zika virus RNA in human and animal specimens by a novel, highly sensitive and specific real-time RT-PCR assay targeting the 5'-untranslated region of Zika virus.
abstract: We developed and evaluated five novel real-time RT-PCR assays targeting conserved regions in the 5'-untranslated region (5'-UTR), envelope (E'), non-structural protein 2A (NS2A), NS5 and 3'-UTR of the ZIKV genome.
  
  
  
  The ZIKV-5'-UTR assay exhibited the lowest in vitro limit of detection (5-10 RNA copies/reaction and 3.0 × 10-1plaque-forming units/ml). Compared to the modified version of a widely adopted RT-PCR assay targeting the ZIKV-E gene, the ZIKV-5'-UTR assay showed better sensitivity in human clinical specimens, and representative mouse specimens, including many organs which are known to be involved in human ZIKV infection but difficult to obtain in clinical settings. The ZIKV-5'-UTR assay detected ZIKV RNA in 84/84 (100.0%) ZIKV-E'-positive and an additional 30/296 (10.1%, P < 0.01) ZIKV-E'-negative mouse specimens. The higher sensitivity of the ZIKV-5'-UTR assay was most significant in kidney and testis/epididymis specimens (P < 0.01). No in vitro or in vivo cross-reactivity was found between the ZIKV-5'-UTR assay and dengue virus, yellow fever virus, Japanese encephalitis virus, West Nile virus, hepatitis C virus and Chikungunya virus.
  
  
  
  The highly sensitive and specific ZIKV-5'-UTR assay may help to improve the laboratory diagnosis of ZIKV infection.
authors.affiliation: State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong, China.|Department of Microbiology, The University of Hong Kong, Hong Kong, China.|Research Centre of Infection and Immunology, The University of Hong Kong, Hong Kong, China.|Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong, China.
authors.forename: Jasper Fuk-Woo
authors.initials: JF
authors.lastname: Chan
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Cyril Chik-Yan
authors.initials: CC
authors.lastname: Yip
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Kah-Meng
authors.initials: KM
authors.lastname: Tee
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Jessica Oi-Ling
authors.initials: JO
authors.lastname: Tsang
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Kenn Ka-Heng
authors.initials: KK
authors.lastname: Chik
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Terance Gi-Wai
authors.initials: TG
authors.lastname: Tsang
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Chris Chung-Sing
authors.initials: CC
authors.lastname: Chan
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Vincent Kwok-Man
authors.initials: VK
authors.lastname: Poon
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Siddharth
authors.initials: S
authors.lastname: Sridhar
authors.affiliation: Key Laboratory of Tropical Diseases and Translational Medicine, Hainan Medical University, Haikou, China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Yin
authors.affiliation: Research Centre of Infection and Immunology, The University of Hong Kong, Hong Kong, China.|Department of Medicine, The University of Hong Kong, Hong Kong, China.
authors.forename: Ivan Fan-Ngai
authors.initials: IF
authors.lastname: Hung
authors.affiliation: Department of Pathology, United Christian Hospital, Hong Kong, China.
authors.forename: Sandy Ka-Yee
authors.initials: SK
authors.lastname: Chau
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Anna Jinxia
authors.initials: AJ
authors.lastname: Zhang
authors.affiliation: Department of Microbiology, The University of Hong Kong, Hong Kong, China.
authors.forename: Kwok-Hung
authors.initials: KH
authors.lastname: Chan
authors.affiliation: State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong, China.|Department of Microbiology, The University of Hong Kong, Hong Kong, China.|Research Centre of Infection and Immunology, The University of Hong Kong, Hong Kong, China.|Carol Yu Centre for Infection, The University of Hong Kong, Hong Kong, China.|Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The University of Hong Kong, Hong Kong, China.
authors.forename: Kwok-Yung
authors.initials: KY
authors.lastname: Yuen
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D003429:Cross Reactions; D006801:Humans; D051379:Mice; D012367:RNA, Viral / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D000071244:Zika Virus / Q000235:genetics*; D000071243:Zika Virus Infection / Q000175:diagnosis* / Q000821:virology
chemical_list: D020121:5' Untranslated Regions; D012367:RNA, Viral
keywords: 
  PCR
  ; Zika; diagnostics; diagnóstico; flavivirus; non traduit; polymerase chain reaction; sin transcribir; untranslated; virus