Monoclonal antibody-based immunohistochemical detection of porcine epidemic diarrhea virus antigen in formalin-fixed, paraffin-embedded intestinal tissues
Monoclonal antibody-based immunohistochemical detection of porcine epidemic diarrhea virus antigen in formalin-fixed, paraffin-embedded intestinal tissues
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DOI：
10.1177/104063879901100512
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发表时间：
1999-09-01
影响因子：
1.5
通讯作者：
Kweon, CH
中科院分区：
文献类型：
--
作者：
Kim, O;Chae, C;Kweon, CH
建立了猪流行性腹泻病毒(PEDV)的免疫组织化学诊断技术。该技术在自然感染PEDV的福尔马林固定、石蜡包埋的仔猪肠道组织上进行了试验，并检测了5种不同的单抗。从自然感染PEDV的仔猪的PLP(4%多聚甲醛、100mM L赖氨酸二盐酸盐、10 mM间高碘酸钠)固定的PEDV感染的Vero细胞或福尔马林固定的石蜡包埋的肠道组织中，一致地检测到PEDV抗原。C9-2-2单抗的反应性最强，背景染色最少，在10头感染猪中检出10头。阳性反应为胞浆阳性。阳性肠细胞分布于空肠和回肠末端及萎缩或融合绒毛的两侧。隐窝内未检测到阳性细胞。盲肠和结肠未见染色。C9-2-2单抗与未感染仔猪、传染性胃肠道病毒(TGEV)和轮状病毒感染仔猪的组织切片反应均未见阳性细胞。选定的抗PEDV单抗在福尔马林固定的石蜡包埋组织切片上测试，在病毒分离不可用时有助于诊断。这种方法在PEDV和TGEV都是流行性传染病的国家特别有价值，并将有助于区分PEDV和TGEV感染。
An immunohistochemistry technique was developed for the diagnosis of porcine epidemic diarrhea virus (PEDV). The technique was tested on formalin-fixed, paraffin-embedded intestinal tissues from piglets naturally infected with PEDV, Five different monoclonal antibodies (MAbs) were tested in this study. PEDV antigen was consistently detected in the PLP (4% paraformaldehyde, 100 mM L-lysine dihydrochloride, 10 mM sodium m-periodate in phosphate-buffered saline)-fixed PEDV-infected Vero cells or formalin-fixed, paraffin-embedded intestinal tissues from piglets naturally infected with PEDV. The C9-2-2 MAb gave the strongest reactivity and least background staining, detecting 10 of 10 infected pigs. The positive reaction was cytoplasmic. Positive enterocytes were distributed over the tip and along the sides of atrophied or fused villi in the jejunum and ileum. Positive-staining cells were not detected in the crypts. No staining was observed in cecum and colon. No positive cells were observed when the C9-2-2 MAb was reacted with the tissue sections from noninfected piglets or from transmissible,gastroenteritus virus (TGEV)- and rotavirus-infected piglets. The selected anti-PEDV MAbs tested on formalin-fixed, paraffin-embedded tissue sections are useful for diagnosis when virus isolation is not available. This method would be of particular value in countries where both PEDV and TGEV are epizootic and would aid in differentiating between PEDV and TGEV infection.