title: Detection, differentiation, and VP1 sequencing of duck hepatitis A virus type 1 and type 3 by a 1-step duplex reverse-transcription PCR assay.
abstract: Duck hepatitis A virus (DHAV) is an infectious pathogen causing fatal duck viral hepatitis in ducklings. Although both the inactivated vaccines and live attenuated vaccines have been used to protect ducklings, DHAV-1 and DHAV-3 still cause significant serious damage to the duck industry in China and South Korea. For rapid detection, differentiation, and epidemic investigation of DHAV in China, a genotype-specific 1-step duplex reverse-transcription (RT) PCR assay was established in this study. The sensitivity and specificity of the developed RT-PCR assay was evaluated with nucleic acids extracted from 2 DHAV reference strains, and 9 other infectious viruses and bacteria. The genotype-specific primers amplified different size DNA fragments encompassing the complete VP1 gene of the DHAV-1 or DHAV-3. The assay detected the liver samples collected from experimentally infected ducklings and dead ducklings collected from different regions of China. Sequence analysis of these DNA fragments indicated that VP1 sequences of DHAV-1 can be used to distinguish wild type and vaccine strains. The phylogenetic analysis of VP1 sequences indicated that the developed RT-PCR assay can be used for epidemic investigation of DHAV-1 and DHAV-3. The developed RT-PCR assay can be used as a specific molecular tool for simultaneous detection, differentiation, and sequencing the VP1 gene of DHAV-1 and DHAV-3, which can be used for understanding the epidemiology and evolution of DHAV.
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China.
authors.forename: X J
authors.initials: XJ
authors.lastname: Wen
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, 46# Xinkang Road, Ya'an, Sichuan 625014, P. R. China chenganchun@vip.163.com.
authors.forename: A C
authors.initials: AC
authors.lastname: Cheng
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, 46# Xinkang Road, Ya'an, Sichuan 625014, P. R. China.
authors.forename: M S
authors.initials: MS
authors.lastname: Wang
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, 46# Xinkang Road, Ya'an, Sichuan 625014, P. R. China.
authors.forename: R Y
authors.initials: RY
authors.lastname: Jia
authors.affiliation: Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, 46# Xinkang Road, Ya'an, Sichuan 625014, P. R. China.
authors.forename: D K
authors.initials: DK
authors.lastname: Zhu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, 46# Xinkang Road, Ya'an, Sichuan 625014, P. R. China.
authors.forename: S
authors.initials: S
authors.lastname: Chen
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, 46# Xinkang Road, Ya'an, Sichuan 625014, P. R. China.
authors.forename: M F
authors.initials: MF
authors.lastname: Liu
authors.affiliation: Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China.
authors.forename: F
authors.initials: F
authors.lastname: Liu
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, P. R. China.
authors.forename: X Y
authors.initials: XY
authors.lastname: Chen
mesh_terms: D000818:Animals; D004372:Ducks; D006506:Hepatitis A / Q000662:veterinary* / Q000821:virology; D030041:Hepatitis A virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D011201:Poultry Diseases / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000662:veterinary; D015678:Viral Structural Proteins / Q000235:genetics* / Q000378:metabolism; D014773:Virology / Q000379:methods*
chemical_list: C096539:VP1 protein, hepatitis A virus; D015678:Viral Structural Proteins
keywords: VP1; duck hepatitis A virus type 1; duck hepatitis A virus type 3; duck viral hepatitis; epidemic investigation