title: Simultaneous detection of severe acute respiratory syndrome, Middle East respiratory syndrome, and related bat coronaviruses by real-time reverse transcription PCR.
abstract: Since severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) coronaviruses (CoVs) share similar characteristics with respect to clinical signs, etiology, and transmission, methods for a rapid and accurate differential diagnosis are important. Therefore, the aim of this study was to develop a duplex real-time reverse transcription (RT)-PCR method for the simultaneous detection of these viruses. Primers and probes that target the conserved spike S2 region of human SARS-CoV, MERS-CoV, and their related bat CoVs were designed. The results of real-time RT-PCR showed specific reactions for each virus with adequate detection limits of 50-100 copies/mL and 5-100 copies/mL using pUC57-SARS-pS2 (a template for SARS-CoV) and pGEM-MERS-S2 (a template for MERS-CoV), respectively. In addition, this real-time RT-PCR system was able to detect the target viruses SARS-like bat CoV and MERS-CoV in bat fecal samples and sputum of MERS patients, respectively. Therefore, this newly developed real-time RT-PCR method is expected to detect not only SARS-CoV and MERS-CoV in humans but also several bat CoVs that are closely related to these viruses in bats.
authors.affiliation: Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea.|College of Veterinary Medicine, Chungbuk National University, Cheongju, 28644, Republic of Korea.
authors.forename: Ji Yeong
authors.initials: JY
authors.lastname: Noh
authors.affiliation: Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea.|Bio-Analytical Science Division, Korea University of Science and Technology (UST), Daejeon, Republic of Korea.
authors.forename: Sun-Woo
authors.initials: SW
authors.lastname: Yoon
authors.affiliation: Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea.
authors.forename: Doo-Jin
authors.initials: DJ
authors.lastname: Kim
authors.affiliation: Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea.|Bio-Analytical Science Division, Korea University of Science and Technology (UST), Daejeon, Republic of Korea.
authors.forename: Moo-Seung
authors.initials: MS
authors.lastname: Lee
authors.affiliation: Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea.
authors.forename: Ji-Hyung
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Pharmacy, College of Pharmacy, Korea University, Sejong, Republic of Korea.
authors.forename: Woonsung
authors.initials: W
authors.lastname: Na
authors.affiliation: Department of Pharmacy, College of Pharmacy, Korea University, Sejong, Republic of Korea.
authors.forename: Daesub
authors.initials: D
authors.lastname: Song
authors.affiliation: Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea. dgjeong@kribb.re.kr.|Bio-Analytical Science Division, Korea University of Science and Technology (UST), Daejeon, Republic of Korea. dgjeong@kribb.re.kr.
authors.forename: Dae Gwin
authors.initials: DG
authors.lastname: Jeong
authors.affiliation: Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea. khk1329@kribb.re.kr.
authors.forename: Hye Kwon
authors.initials: HK
authors.lastname: Kim
mesh_terms: D000818:Animals; D002685:Chiroptera / Q000821:virology; D003332:Coronaviridae / Q000235:genetics / Q000302:isolation & purification*; D018352:Coronavirus Infections / Q000821:virology; D003937:Diagnosis, Differential; D005243:Feces / Q000821:virology; D006801:Humans; D057230:Limit of Detection; D065207:Middle East Respiratory Syndrome Coronavirus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000302:isolation & purification*; D016415:Sequence Alignment; D045169:Severe Acute Respiratory Syndrome / Q000821:virology; D013183:Sputum / Q000821:virology
chemical_list: D012367:RNA, Viral