Wen X J, Cheng A C, Wang M S, Jia R Y, Zhu D K, Chen S, Liu M F, Liu F, Chen X Y
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.
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
Poult Sci. 2014 Sep;93(9):2184-92. doi: 10.3382/ps.2014-04024. Epub 2014 Jul 10.
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.
鸭甲型肝炎病毒（DHAV）是一种可导致雏鸭致命性鸭病毒性肝炎的传染性病原体。尽管灭活疫苗和减毒活疫苗均已用于保护雏鸭，但DHAV - 1和DHAV - 3仍在中国和韩国给养鸭业造成重大严重损失。为了在中国快速检测、鉴别DHAV并进行流行病学调查，本研究建立了一种基因型特异性一步双重逆转录（RT）PCR检测方法。利用从2株DHAV参考毒株以及9种其他传染性病毒和细菌中提取的核酸，对所建立的RT - PCR检测方法的敏感性和特异性进行了评估。基因型特异性引物扩增出了包含DHAV - 1或DHAV - 3完整VP1基因的不同大小的DNA片段。该检测方法检测了从实验感染雏鸭以及从中国不同地区采集的死亡雏鸭身上获取的肝脏样本。对这些DNA片段的序列分析表明，DHAV - 1的VP1序列可用于区分野生型和疫苗株。对VP1序列的系统发育分析表明，所建立的RT - PCR检测方法可用于DHAV - 1和DHAV - 3的流行病学调查。所建立的RT - PCR检测方法可作为一种特异性分子工具，用于同时检测、鉴别DHAV - 1和DHAV - 3的VP1基因并对其进行测序，这可用于了解DHAV的流行病学和进化情况。
