石斑鱼蛙病毒在鞍带石斑鱼中的组织分布

摘要

Grouper ranavirus infected many grouper species and caused severe mortality. We isolated a grouper iridovirus( genus Ranaυirus)named GIV-R-SY1301 in Sanya by using cell culture,to study the tissue distribution of GIV-R-SY1301 in experimentally infected farmed Epinephelus lanceolatus by in situ hybridization and real-time PCR. The GIV-R-SY1301 was widespread in all tissues from infected fish. Strong hybridization signals were obtained from kidney and spleen,while intermediate intensity signals were ob-served in stomach,gill and liver. The weakest signals were obtained from heart and no signal was found in brain and eye. The real-time PCR results show that there were most viruses copies in spleen,kidney and heart(4. 5 × 104 ~ 6. 85 × 107 copies·μg-1 )DNA in different tissues,which was similar with the result of in situ hybridization.%石斑鱼蛙病毒能够感染多种石斑鱼并造成严重死亡。研究利用原位杂交和荧光定量 PCR 技术对蛙病毒三亚株 GIV-R-SY1301在人工感染发病的鞍带石斑鱼（Epinephelus lanceolatus）不同组织中的分布进行定位、定量研究。原位杂交结果显示 GIV-R-SY1301广泛存在于发病鱼的多个组织中，脾脏和肾脏中杂交信号最强，其次是肠道、胃、鳃、肝，心脏中也有少量的杂交信号，而脑和眼组织中没有杂交信号存在。荧光定量 PCR 检测结果显示 GIV-R-SY1301在不同组织中的病毒含量为4.5×104~6.85×107拷贝·μg -1 DNA，其中脾脏、心脏和肾脏中最多。组织原位杂交和荧光定量 PCR 结果基本一致。