title: Development of RT-LAMP and real-time RT-PCR assays for the rapid detection of the new duck Tembusu-like BYD virus.
abstract: A new duck Tembusu virus (TMUV), also known as BYD virus, has been identified as the causative agent for the emerging duck egg-drop syndrome in mainland China. The rapid spread and wide distribution of the new TMUV in mainland China result in heavy loss to the poultry industry and pose great threats to public health. Rapid and sensitive detection methods are critical for prevention and control of TMUV infections. In this study, a reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) and an SYBR Green-I-based real-time RT-PCR assay specific for the duck TMUV were developed and validated with laboratory and field samples, respectively. The detection limits were 1 × 10(-4) and 1 × 10(-3) PFU per reaction for the RT-LAMP assay and real-time RT-PCR assay, respectively. The specificities were analyzed with other related members of the genus Flavivirus, and no cross-reaction was observed. Furthermore, both assays were evaluated with field samples, and they exhibited high sensitivity and specificity. In addition, the real-time RT-PCR assay worked well in viral load analysis, which revealed that the spleen may be the primary target for the replication of new duck TMUV in ducks. The TMUV-specific RT-LAMP and real-time RT-PCR assays will provide useful tools for the diagnosis and epidemiological surveillance of TMUV infection.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Department of Virology, Beijing Institute of Microbiology and Epidemiology, Fengtai District, 100071 Beijing, China. jiang_tao@126.com
authors.forename: Tao
authors.initials: T
authors.lastname: Jiang
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.forename: Yong-Qiang
authors.initials: YQ
authors.lastname: Deng
authors.forename: Jing-Liang
authors.initials: JL
authors.lastname: Su
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Xu
authors.forename: Zhi-Hui
authors.initials: ZH
authors.lastname: Liu
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Li
authors.forename: Xue-Dong
authors.initials: XD
authors.lastname: Yu
authors.forename: Shun-Ya
authors.initials: SY
authors.lastname: Zhu
authors.forename: George Fu
authors.initials: GF
authors.lastname: Gao
authors.forename: E-De
authors.initials: ED
authors.lastname: Qin
authors.forename: Cheng-Feng
authors.initials: CF
authors.lastname: Qin
mesh_terms: D000818:Animals; D001921:Brain / Q000821:virology; D002681:China / Q000453:epidemiology; D004372:Ducks; D005416:Flavivirus / Q000235:genetics / Q000302:isolation & purification*; D018177:Flavivirus Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D008099:Liver / Q000821:virology; D011201:Poultry Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013577:Syndrome; D019562:Viral Load