Chvala S, Fragner K, Hackl R, Hess M, Weissenböck H
Department of Pathobiology, Institute of Pathology and Forensic Veterinary Medicine, Veterinärplatz 1, A-1210 Vienna, Austria.
J Comp Pathol. 2006 Feb-Apr;134(2-3):211-8. doi: 10.1016/j.jcpa.2005.11.002.
An in-situ hybridization (ISH) procedure was developed for the detection of Cryptosporidium sp. in paraffin wax-embedded tissues with a digoxigenin-labelled probe targeting the 18S rRNA. This technique was used in addition to traditional methods, such as haematoxylin and eosin staining, periodic acid-Schiff reaction, transmission electron microscopy and the polymerase chain reaction, to examine the bursa of Fabricius (BF), conjunctiva and other tissues from 20 domestic geese aged 16-36 days for the presence of cryptosporidia. Positive signals were found to a moderate or marked extent in both conjunctival samples (89%) and BF samples (88%) but not in other tissues. Sequencing of the PCR amplification product revealed identity with Cryptosporidium baileyi. The infected geese showed no clinical signs and only scanty histological lesions. These results confirm reports showing that young waterfowl are especially vulnerable to cryptosporidium infection and indicate that the BF and conjunctiva are the preferred sites for the presence of the protozoon. ISH proved a good method for detecting and identifying even small numbers of cryptosporidia in tissue sections.
开发了一种原位杂交（ISH）程序，用于检测石蜡包埋组织中的隐孢子虫属，使用地高辛标记的针对18S rRNA的探针。除了苏木精和伊红染色、过碘酸-希夫反应、透射电子显微镜和聚合酶链反应等传统方法外，该技术还用于检查20只16至36日龄家鹅的法氏囊（BF）、结膜和其他组织中是否存在隐孢子虫。在结膜样本（89%）和BF样本（88%）中均发现了中度或明显程度的阳性信号，但在其他组织中未发现。PCR扩增产物的测序显示与贝利隐孢子虫一致。受感染的鹅没有临床症状，只有少量组织学病变。这些结果证实了有关幼龄水禽特别易受隐孢子虫感染的报道，并表明BF和结膜是该原生动物存在的首选部位。ISH被证明是一种检测和识别组织切片中即使少量隐孢子虫的好方法。
