Huang Canhua, Zhang Xiaobo, Gin Karina Y H, Qin Qi Wei
Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260, Singapore.
J Virol Methods. 2004 May;117(2):123-8. doi: 10.1016/j.jviromet.2004.01.002.
A DNA probe of 531 base pairs for Singapore grouper iridovirus (SGIV) was generated by polymerase chain reaction and labeled with nonradioactive digoxigenin. An in situ hybridization based method was developed to detect SGIV in formalin-fixed tissues from maricultured Malabar grouper, Epinephelus malabaricus Bloch and Schneider. The in situ hybridization detected SGIV in the kidney, spleen, liver, intestine, stomach and gills from naturally infected fish. Strong hybridization signals were obtained from the kidney and spleen tissues, while intermediate intensity signals were observed in the intestine and liver tissues. The weakest signals were obtained from the stomach and gills. The signals were located specifically within epithelial, endothelial and sub-endothelial hypertrophic cells in all tested tissues. The in situ hybridization procedure will provide an important diagnostic tool to complement histopathological methods, and contribute to epidemiological studies on the origin and distribution of iridovirus in mariculture.
通过聚合酶链反应生成了一段531个碱基对的新加坡石斑鱼虹彩病毒（SGIV）DNA探针，并用非放射性地高辛配基进行标记。开发了一种基于原位杂交的方法，用于检测养殖的斜带石斑鱼（Epinephelus malabaricus Bloch and Schneider）福尔马林固定组织中的SGIV。原位杂交在自然感染鱼的肾脏、脾脏、肝脏、肠道、胃和鳃中检测到了SGIV。在肾脏和脾脏组织中获得了强杂交信号，而在肠道和肝脏组织中观察到中等强度信号。在胃和鳃中获得的信号最弱。在所有测试组织中，信号特异性地位于上皮细胞、内皮细胞和内皮下肥大细胞内。原位杂交程序将提供一种重要的诊断工具，以补充组织病理学方法，并有助于对虹彩病毒在海水养殖中的起源和分布进行流行病学研究。
