title: Development and application of a reverse transcription loop-mediated isothermal amplification method for rapid detection of Duck hepatitis A virus type 1.
abstract: We developed and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting Duck hepatitis A virus type 1 (DHAV-1). The amplification could be finished in 1 h under isothermal conditions at 63 °C by employing a set of four primers targeting the 2C gene of DHAV-1. The RT-LAMP assay showed higher sensitivity than the RT-PCR with a detection limit of 0.1 ELD(50) 0.1 ml(-1) of DHAV-1. The RT-LAMP assay was highly specific; no cross-reactivity was observed from the samples of other related viruses, bacteria, allantoic fluid of normal chicken embryos, or the livers of uninfected ducks. Thirty clinical samples were subjected to detection by RT-LAMP, RT-PCR, and virus isolation, which obtained completely consistent, positive results. As a simple, rapid, and accurate detection method, this RT-LAMP assay has important potential applications in the clinical diagnosis of DHAV-1.
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Limin
authors.initials: L
authors.lastname: Yang
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Yuhai
authors.initials: Y
authors.lastname: Bi
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.forename: Wenjun
authors.initials: W
authors.lastname: Liu
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D002642:Chick Embryo; D003429:Cross Reactions; D017931:DNA Primers / Q000235:genetics; D004372:Ducks / Q000821:virology*; D004371:Hepatitis Virus, Duck / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D006524:Hepatitis, Viral, Animal / Q000175:diagnosis / Q000821:virology; D008099:Liver / Q000821:virology; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014776:Virus Cultivation / Q000379:methods
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D017361:Viral Nonstructural Proteins; C076023:2C protein, viral