Chen Zong-yan, Cheng An-chun, Wang Ming-shu, Xu Da-wei, Jia Renyong, Guo Yu-fei, Zeng Weng
Key Laboratory of Animal Disease and Human Health of Sichuan Province, Yaan, China.
Avian Pathol. 2009 Apr;38(2):129-34. doi: 10.1080/03079450902737862.
In order to define clearly the conditions leading to the outcome of acute duck hepatitis B virus (DHBV) infection, 1-day-old Pekin ducklings were infected with DHBV by different routes and given different doses of inoculum. Groups of 24 ducklings were inoculated either intravenously via the vena cruralis, or intraperitoneally with pooled serum containing either 1.6 x 10(7) or 1.6 x 10(4) DHBV genomes. One control duck from each group was inoculated with an equal volume of normal duck serum. A sensitive and reproducible real-time polymerase chain reaction assay based on TaqMan technology was developed for the detection and quantitation of DHBV DNA in the serum and liver. DHBAg was observed in the hepatocytes by immunohistochemistry. Histological changes in the liver tissue were also observed. The results demonstrate that ducklings at each time point and in all groups developed detectable viraemia. In each group, DHBV DNA in the liver was at a lower level than in serum and the peak DNA titre was found in serum earlier than in the liver. In the low-dose groups it was always at a lower level than in the high-dose groups. The DHBV replication levels appeared to be directly related to the number of DHBAg-positive hepatocytes. The variation trends of DHBAg-positive hepatocytes were similar in the high-dose groups. Histological changes were associated with liver viral DNA levels. We suggest that this dose and route of inoculation can be used as a model to study acute DHBV infections.
为了明确导致急性鸭乙型肝炎病毒（DHBV）感染结果的条件，对1日龄北京鸭雏通过不同途径感染DHBV，并给予不同剂量的接种物。将24只鸭雏分为几组，分别通过胫静脉进行静脉接种，或通过腹腔注射含有1.6×10⁷或1.6×10⁴个DHBV基因组的混合血清。每组取1只对照鸭接种等量的正常鸭血清。开发了一种基于TaqMan技术的灵敏且可重复的实时聚合酶链反应检测方法，用于检测和定量血清及肝脏中的DHBV DNA。通过免疫组织化学在肝细胞中观察到DHBAg。还观察了肝组织的组织学变化。结果表明，各时间点和所有组的鸭雏均出现了可检测到的病毒血症。在每组中，肝脏中的DHBV DNA水平低于血清中的水平，且血清中DNA滴度峰值出现的时间早于肝脏。在低剂量组中，其水平始终低于高剂量组。DHBV复制水平似乎与DHBAg阳性肝细胞的数量直接相关。高剂量组中DHBAg阳性肝细胞的变化趋势相似。组织学变化与肝脏病毒DNA水平相关。我们认为这种接种剂量和途径可作为研究急性DHBV感染的模型。
