Kim B, Chae C
Department of Veterinary Pathology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Kyounggi-Do, Republic of Korea.
Can J Vet Res. 2001 Jan;65(1):33-7.
Archived formalin-fixed, paraffin-embedded tissues from 25 pigs naturally infected with transmissible gastroenteritis virus (TGEV) were examined by in situ hybridization for TGEV nucleic acid using a nonradioactive digoxigenin-labeled cDNA probe that targeted the nucleocapsid sequence of TGEV strains. The results of in situ hybridization for the detection of TGEV were compared with virus isolation (VI), a fluorescent antibody test (FAT), and transmission electron microscopy (TEM). VI, FAT, and TEM were tested over a course of time before the in situ hybridization was performed. Positive hybridization signals were detected in duodenal, jejunal, and ileal enterocytes from 21 pigs. Hybridization signals were confined to the cytoplasm. Intestinal specimens from 25 piglets were evaluated by 4 tests. Twenty-one of 25 were positive by in situ hybridization. Of these 21 samples, 5 (24%) were positive for TGEV by all 4 tests, 15 (71%) were positive by FAT, 14 (67%) were positive by VI, and 6 (29%) were positive by TEM. In situ hybridization for the detection of TGEV in formalin-fixed, paraffin-embedded tissues provides a rapid means of confirmation of a histopathological diagnosis of TGEV without virus isolation, or when only formalin-fixed intestinal specimens were available.
对25头自然感染传染性胃肠炎病毒(TGEV)的猪的存档福尔马林固定石蜡包埋组织进行了原位杂交检测,使用针对TGEV毒株核衣壳序列的非放射性地高辛标记cDNA探针检测TGEV核酸。将检测TGEV的原位杂交结果与病毒分离(VI)、荧光抗体试验(FAT)和透射电子显微镜(TEM)进行了比较。在进行原位杂交之前的一段时间内对VI、FAT和TEM进行了检测。在21头猪的十二指肠、空肠和回肠肠细胞中检测到阳性杂交信号。杂交信号局限于细胞质。通过4种检测方法对25头仔猪的肠道标本进行了评估。25头中有21头通过原位杂交呈阳性。在这21个样本中,5个(24%)通过所有4种检测方法均为TGEV阳性,15个(71%)通过FAT呈阳性,14个(67%)通过VI呈阳性,6个(29%)通过TEM呈阳性。在福尔马林固定石蜡包埋组织中检测TGEV的原位杂交提供了一种快速确认TGEV组织病理学诊断的方法,无需病毒分离,或在仅有福尔马林固定的肠道标本时使用。