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应用显色原位杂交法检测猫肠道组织标本中的胎儿三毛滴虫和五毛滴虫。

Detection of Tritrichomonas foetus and Pentatrichomonas hominis in intestinal tissue specimens of cats by chromogenic in situ hybridization.

机构信息

Institute of Pathology and Forensic Veterinary Medicine, Department of Pathobiology, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria.

出版信息

Vet Parasitol. 2012 Feb 10;183(3-4):209-14. doi: 10.1016/j.vetpar.2011.07.050. Epub 2011 Aug 4.

Abstract

In this retrospective study 102 cats were analyzed for the presence of trichomonads in intestinal tissue sections using chromogenic in situ hybridization (CISH). Two intestinal trichomonad species are described in cats: Pentatrichomonas hominis and Tritrichomonas foetus. While P. hominis is considered a mere commensal, T. foetus has been found to be the causative agent of feline large-bowel diarrhea. For the detection of both agents within intestinal tissue CISH assays using three different probes were performed. In the first CISH run a probe specific for all relevant members of the order Trichomonadida (OT probe) was used. In a second CISH run all positive samples were further examined on three consecutive tissue sections using the OT probe, a probe specific for the family of Tritrichomonadidae (Tritri probe) and a newly designed probe specifically detecting P. hominis (Penta hom probe). In total, four of the 102 cats were found to be positive with the OT probe. Thereof, one cat gave a positive reaction with the P. hominis probe and three cats were positive with the T. foetus probe. All Trichomonas-positive cats were pure-bred and between 8 and 32 weeks of age. In one cat positive for T. foetus large amounts of parasites were found in the gut lumen and invading the intestinal mucosa. The species of the detected trichomonads were confirmed by polymerase chain reaction and nucleotide sequencing of a part of the 18S ribosomal RNA gene. In this study, the usefulness of CISH to detect intestinal trichomonads within feline tissue samples was shown. Additionally, the specific detection of P. hominis using CISH was established. Generally, it was shown that CISH is well suited for detection and differentiation of trichomonosis in retrospective studies using tissue samples.

摘要

在这项回顾性研究中,使用显色原位杂交(CISH)分析了 102 例猫肠道组织切片中是否存在滴虫。猫体内有两种肠滴虫:五毛滴虫和胎儿滴虫。虽然五毛滴虫被认为只是共生菌,但胎儿滴虫已被发现是猫大肠性腹泻的病原体。为了在肠道组织中检测这两种病原体,使用了三种不同的探针进行 CISH 检测。在第一次 CISH 运行中,使用了一种针对 Trichomonadida 目所有相关成员的探针(OT 探针)。在第二次 CISH 运行中,所有阳性样本在三个连续的组织切片上使用 OT 探针、一种针对 Tritrichomonadidae 科的探针(Tritri 探针)和一种新设计的专门检测五毛滴虫的探针(Penta hom 探针)进行了连续检测。总共,在 102 只猫中有 4 只被 OT 探针检测为阳性。其中,一只猫的 P. hominis 探针呈阳性反应,三只猫的 T. foetus 探针呈阳性反应。所有 Trichomonas 阳性猫均为纯种,年龄在 8 至 32 周之间。在一只 T. foetus 阳性猫的肠道腔和肠道黏膜中发现了大量寄生虫。通过聚合酶链反应和 18S 核糖体 RNA 基因的一部分核苷酸测序,确认了检测到的滴虫的种类。在这项研究中,展示了 CISH 用于检测猫肠道组织样本中肠道滴虫的有用性。此外,还建立了使用 CISH 特异性检测五毛滴虫的方法。一般来说,研究表明 CISH 非常适合使用组织样本进行回顾性研究中滴虫病的检测和鉴别。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4be/3253383/ffeefc143349/gr1.jpg

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