Singh Amarjit, Weissenböck Herbert, Hess Michael
Clinic for Avian, Reptile and Fish Medicine, Department for Farm Animals and Herd Management, University of Veterinary Medicine, Veterinärplatz 1, A-1210 Vienna, Austria.
Exp Parasitol. 2008 Apr;118(4):505-13. doi: 10.1016/j.exppara.2007.11.004. Epub 2007 Nov 17.
In the present investigation, a polyclonal antibody-based immunohistochemical technique was developed to localize Histomonas meleagridis in formalin-fixed, paraffin-embedded tissues of experimentally infected turkeys. The developed technique was highly specific for histomonads as no immunohistochemical reaction was observed with cultures of Tetratrichomonas gallinarum, Trichomonas gallinae and Blastocystis sp. In addition, tissues positive for various other protozoan parasites and fungi were also tested to evaluate the specificity of the technique. It was possible to detect immunohistochemically histomonad antigens in all the tested samples (n=5) of caecum, liver, spleen and lung from infected turkeys, 3 out of 5 bursa of Fabricius, 1 out of 2 bone marrow, 2 out of 5 heart and 1 out of 5 each of proventriculus, pancreas and cerebellum. An immunohistochemical reaction indicative of presence of histomonads was also detected in blood vessels of various organs that indicated a possible hematogenous route of spread of the parasite in the host. A comparative study with routine diagnostic staining techniques indicated a high sensitivity and specificity of the newly developed immunohistochemical technique. Altogether, the technique developed can be used to study the sequential pathogenesis of histomonosis in turkeys and to obtain new insights into the mechanisms of interaction with the host tissues.
在本研究中,开发了一种基于多克隆抗体的免疫组织化学技术,用于在实验感染火鸡的福尔马林固定、石蜡包埋组织中定位火鸡组织滴虫。所开发的技术对组织滴虫具有高度特异性,因为用鸡四毛滴虫、禽毛滴虫和芽囊原虫的培养物未观察到免疫组织化学反应。此外,还对各种其他原生动物寄生虫和真菌呈阳性的组织进行了检测,以评估该技术的特异性。在感染火鸡的盲肠、肝脏、脾脏和肺的所有测试样本(n = 5)、5个法氏囊中3个、2个骨髓中1个、5个心脏中2个以及腺胃、胰腺和小脑各5个中的1个中,通过免疫组织化学方法检测到了组织滴虫抗原。在各个器官的血管中也检测到了表明存在组织滴虫的免疫组织化学反应,这表明寄生虫在宿主体内可能存在血源性传播途径。与常规诊断染色技术的比较研究表明,新开发的免疫组织化学技术具有高灵敏度和特异性。总之,所开发的技术可用于研究火鸡组织滴虫病的序贯发病机制,并获得与宿主组织相互作用机制的新见解。