中国对虾肝胰腺细小病毒感染的免疫病理学研究  

AN IMMUNE PATHOLOGICAL STUDY ON THE HEPATOPANCREATIC PARVOVIRUS INFECTION OF PENAEUS CHINENSIS

作　　者：薛清刚[1] 钱鸣亮[2] 罗挽涛[1] 宋庆云[1] 杨虹[1] 王文兴[1] 

机构地区：[1]国家海洋局第一海洋研究所,青岛266003 [2]湛江海洋大学水产学院养殖系,湛江524000

出　　处：《海洋与湖沼》1998年第1期9-14,T001,共7页Oceanologia Et Limnologia Sinica

基　　金：国家自然科学基金!39000083;国家攀登计划B!PDB-6-6-2

摘　　要：于1992－1993年，用兔抚对虾肝胰腺细小病毒IgG和辣根过氧化物酶标记的羊抗兔IgG建立免疫组织病理检测技术，并用来对肝胰腺细小病毒感染的中国对虾肝胰腺组织作病理学分析。该方法利用抗原－抗体之间特异性结合的特点，首先令兔抗肝胰腺细小病毒抗体与相应的病毒成分结合，然后以酶标记的羊抗兔免疫球蛋白抗体与巴结合到病毒上的抗体作用，并通过酶显色反应，揭示病毒和相应病变结构的存在。结果显示，免疫酶染色技术将病毒包涵体和因病毒感染而肿胀的细胞核染成金黄至棕色；病毒可游离存在于细胞核腔和核膜内面；完整的包涵体常脱落到肝胰腺管管腔内。从而提示，中国对虾肝胰腺细小病毒多以脱落包涵体的形式造成水平传播；从细胞病理角度，细胞病变应分为细胞核肿胀的早期、包涵体形成的中期和细胞破裂释放包涵体的后期三个阶段。研究结果从一个侧面证实了肝胰腺细小病毒的致病性特征。During 1992- 1993, hepatopancreatic pareovirus (HPV) infected hepatopancreas ofPenaeus chinensis was histopathologically studied using the method of immun enzyme stalning. Withthe rabbit anh-hepatoPanCreatic Pareovirus IgG (PV-IgG) which was prepared in labothery byimmedzing rabbtS with Purified HPV and commercially available horsendsh Peroxidase labeledsheep anti-rabhit IgG (HRP-IgG), an indirect immune enzyme staining procedure for detechng thehepatopanreatic parvovirus in a sechon of Penaeus chinensis hepatopancreas was estabished. Thehepatoparereas from the diseased Penaeus chinensis was fixed in Davison's solution and paraffinsechons of about 5μm were routinely prepared. After treaiment with 0.5% H2O2 and 3% bovineserum albon, the sechon was incubated with PV-IgG at 4t ovendght and, following washingwith PBS, incubthed with HRP-IgG for 60min at room temperabe- Following colohoon with 3'3-diaminobenzidine (DAB), the samples were examined under light microscope. With this procedure,the nuclei of hepatopancreatic epithelium of Penaeus chinensis tha were infected by thehepatopancreatic parvovirus were strined to be yellow or brown in color. Both virus inclusion body(Plate I: 1, IB) and some inclusion body free but enlarged nuclei (Plate 1:3, N) can bespecifically stalned. The cytoplasm of the host cell and the nuclei of noninfected epithelium werenot stajned. Besides the inclusion bod, the cavity (Plate 1:2, VS) between nucleus membran andinclusion body and the membran (Plate I:3, NM) of the inclusion bod-containing nuclei werestained strongly, indicahng tha some virus particles exit freely in the host nucleus. The virusinclusion bodies can leave the host cell (Plate I: 5-6, IB) and drop into hepatopancreatic tubes(Plate I: 7, IB); thes indicates horizontal transmission through the digeshve system. Thecytopathological changes of the infected cells are divided into 3 phases. In Phase I (earycytopathological stage), the nuclei are infected with no inclusion body observed. Phase II (middlestage) is characterized b

关 键 词：肝胰腺细小病毒 中国对虾 免疫病理学 免疫酶 

分 类 号：S945.26[农业科学—水产养殖]