title: Development of a novel dengue virus serotype-specific multiplex real-time reverse transcription-polymerase chain reaction assay for blood screening.
abstract: Dengue fever is caused by four related RNA viruses of the genus Flavivirus, dengue virus (DENV)-1, -2, -3, and -4, which are transmitted to humans by mosquitoes. Although DENV is not endemic in Japan, an autochthonous dengue outbreak occurred in 2014. Several transfusion-transmitted cases have also been reported after the use of blood and plasma products in DENV-endemic countries. The aim of this study was to develop a novel multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay for DENV blood screening.
  
  
  
  Large-scale oligonucleotide screening was performed to obtain DENV-specific primers and probes using a variety of DENV clinical isolates. A multiplex RT-PCR assay was then developed using the identified oligonucleotides and the ability of this assay to detect DENV RNA was evaluated.
  
  
  
  A number of oligonucleotides suitable for DENV RNA detection were identified and a novel DENV serotype-specific multiplex RT-PCR assay was successfully established. Comparative analysis revealed that the multiplex assay could detect levels of viral contamination as low as 100 viral copies/mL.
  
  
  
  This established serotype-specific multiplex RT-PCR assay provides a simple, sensitive, and quantitative detection method for DENV, which could be applied in the screening of blood samples to prevent transfusion-transmitted DENV infection.
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Kenta
authors: K
authors: Tezuka
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Madoka
authors: M
authors: Kuramitsu
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Kazu
authors: K
authors: Okuma
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Kiyoko
authors: K
authors: Nojima
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Kumiko
authors: K
authors: Araki
authors: Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan.
authors: Naoya
authors: N
authors: Shinohara
authors: Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan.
authors: Chieko
authors: C
authors: Matsumoto
authors: Central Blood Institute, Blood Service Headquarters, Japanese Red Cross Society, Tokyo, Japan.
authors: Masahiro
authors: M
authors: Satake
authors: Department of Virology 1, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Tomohiko
authors: T
authors: Takasaki
authors: Department of Virology 1, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Masayuki
authors: M
authors: Saijo
authors: Department of Virology 1, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Ichiro
authors: I
authors: Kurane
authors: Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
authors: Isao
authors: I
authors: Hamaguchi
mesh_terms: D058731:Blood Safety; D003715:Dengue / Q000175:diagnosis* / Q000517:prevention & control / Q000635:transmission; D003716:Dengue Virus / Q000235:genetics*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000097:blood; D060888:Real-Time Polymerase Chain Reaction; D065288:Serogroup; D065227:Transfusion Reaction
chemical_list: D012367:RNA, Viral