title: Comparison of real-time reverse transcription loop-mediated isothermal amplification and real-time reverse transcription polymerase chain reaction for duck Tembusu virus.
abstract: Duck Tembusu virus (DTMUV) has caused huge losses to the poultry industry in China since the spring of 2010. The development of a rapid, convenient, and reliable method to diagnose this emerging duck infectious disease is critical. In the present study, a real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was compared with the real-time reverse transcription polymerase chain reaction (RT-PCR) for detection of DTMUV. The sensitivity of real-time RT-LAMP was equal to that of the real-time RT-PCR, with a detection limit of 0.01 ELD(50) (50% egg lethal dose). The specificity of the real-time RT-LAMP and real-time RT-PCR was confirmed using RNAs and DNAs extracted from related viruses which cause duck infections. The reproducibility of the real-time RT-PCR assay was better than that of the real-time RT-LAMP. Only three results from 96 tissue samples differed between the real-time RT-LAMP and this real-time RT-PCR, confirming the reliability of these methods. This study indicated that the real-time RT-LAMP is simpler, less time-consuming, and more convenient than the real-time RT-PCR. With its high sensitivity, specificity, and convenience, the real-time RT-LAMP is a practical molecular diagnostic method for rapid and quantitative detection of DTMUV infection in a resource-limited setting.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China. Liping.yan912@gmail.com
authors.forename: Liping
authors.initials: L
authors.lastname: Yan
authors.forename: Shan
authors.initials: S
authors.lastname: Peng
authors.forename: Pixi
authors.initials: P
authors.lastname: Yan
authors.forename: Jiewen
authors.initials: J
authors.lastname: Zhou
authors.forename: Qiaoyang
authors.initials: Q
authors.lastname: Teng
authors.forename: Guoxin
authors.initials: G
authors.lastname: Li
authors.forename: Xuesong
authors.initials: X
authors.lastname: Li
authors.forename: Zejun
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D002681:China; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004372:Ducks; D005416:Flavivirus / Q000302:isolation & purification*; D018177:Flavivirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*