College of Veterinary Medicine, Shandong Agricultural University, Taian, 271018, Shandong, China.
Virol Sin. 2012 Aug;27(4):248-53. doi: 10.1007/s12250-012-3255-8. Epub 2012 Jul 28.
To investigate the relationship of the variation of virulence and the external capsid proteins of the pandemic duck hepatitis A virus type 1 (DHAV-1) isolates, the virulence, cross neutralization assays and the complete sequence of the virion protein 1 (VP1) gene of nine virulent DHAV-1 strains, which were isolated from infected ducklings with clinical symptoms in Shandong province of China in 2007-2008, were tested. The fifth generation duck embryo allantoic liquids of the 9 isolates were tested on 12-day-old duck embryos and on 7-day-old ducklings for the median embryonal lethal doses (ELD(50)s) and the median lethal doses (LD(50)s), respectively. The results showed that the ELD(50)s of embryonic duck eggs of the 9 DHAV-1 isolates were between 1.9 × 10(6)/mL to 1.44 × 10(7)/mL, while the LD(50)s were 2.39 × 10(5)/mL to 6.15 × 10(6)/mL. Cross-neutralization tests revealed that the 9 DHAV-1 isolates were completely neutralized by the standard serum and the hyperimmune sera against the 9 DHAV-1 isolates, respectively. Compared with other virulent, moderate virulent, attenuated vaccine and mild strains, the VP1 genes of the 9 strains shared 89.8%-99.7% similarity at the nucleotide level and 92.4%-99.6% at amino acid level with other DHAV-1 strains. There were three hypervariable regions at the C-terminus (aa 158-160, 180-193 and 205-219) and other variable points in VP1 protein, but which didn't cause virulence of DHAV-1 change.
为了研究鸭甲肝病毒 1 型(DHAV-1)流行株毒力变异与其外壳蛋白的关系,对 2007-2008 年山东发病雏鸭分离的 9 株强毒 DHAV-1 进行了致弱、交叉中和试验和 VP1 基因全序列测定。将 9 株 DHAV-1 的第 5 代尿囊液分别对 12 日龄鸭胚和 7 日龄雏鸭进行半数致死量(ELD50)和半数致死剂量(LD50)测定,结果显示,9 株 DHAV-1 的 ELD50 在 1.9×106~1.44×107/mL,LD50在 2.39×105~6.15×106/mL。交叉中和试验结果表明,9 株 DHAV-1 均能被标准阳性血清和自家分离株的高免血清完全中和。与其他强毒、中等毒力、弱毒疫苗株和低毒分离株相比,9 株的 VP1 基因核苷酸序列同源性为 89.8%~99.7%,氨基酸序列同源性为 92.4%~99.6%。VP1 蛋白 C 末端存在 3 个高变区(aa 158-160、180-193、205-219)和其他变点点,但未引起 DHAV-1 毒力发生变化。