猪流行性腹泻的病理学研究——免疫酶组化法(间接法)抗原定位的研究

EXPERIMENTAL STUDY ON THE PATHOLOGY OF PORCINE EPIDEMIC DIARRHOEA-IMMUNOPEROXIDASE STAINING STUDY

本试验给8头3日龄同窝仔猪经口感染猪流行性腹泻病毒“吉”毒株的猪胎肠上皮原代单层细胞培养物,于感染后在第18、30、45、96小时扑杀。以免疫组化法(间接法)进行抗原定位的研究。研究表明:猪流行性腹泻病毒主要侵袭、感染仔猪小肠绒毛上皮细胞,此外还感染大肠粘膜上皮细胞、肠腺上皮细胞、肠系膜淋巴结的巨噬细胞,而与TCE的全身感染不同;以感染30小时,腹泻10小时左右,空肠中、后部阳性细胞最多;腹泻10小时后,小肠绒毛上皮有的变成矮立方形、扁平上皮,有的空泡化,还有的仍为柱状,但这些细胞中都见有阳性反＊，闪而证明这些细胞都畜病毒抗原存在。腹泻1则、时左右的病猪，取其空肠中、后部等病料，用hU切氏液固定、、蜡包埋切片，免疫酶组化法（间接法）染色，可作为流行性腹泻病猪死后诊断的一种特异性方法；石蜡切片以O．15％胰酶在37”C温箱湿盒中消化45分钟最为适宜；为防止脱片，使用乳白胶（工呵OQ）粘片效果最佳；组织切片制备中使用甲苯作为透明剂其免疫酶组化法染色效果优于二甲苯；使用IIRpspA’i呐标免抗猪几G，二行效果无明显差异。

Eight 3-day-old piglets from the same litter were dosed orally with 5 ml PEDV fluids containing 105/0.05 ml.They were killed at 18, 30,45, 96h.after infection.Three piglets were used as controls and killed at 20, 45, 96h after infection.The distribution of 'J' strain of PEDV antigen was found mainly in the epithelial cells covering the small intestinal villi and occasionally in intestinal cryst epithelium and phagocytes in me-sentric lymph nodes. The largest number of positive cells was found in the middle and lower jejunum 10h after diarrhoea. Three type of small intestinal epithelial cells: columnar, cubic and vacuolated cells, appeared 10h after diarrheoa. The viral antigen was also found in all these cells. The method used in this experiment had specificity in diagnosing PED.