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随机扩增多态性DNA分析以及ITS1/5.8S rRNA/ITS2和铁氢化酶测序作为鸡三毛滴虫潜在致病分离株基因分类的工具

RAPD analysis and sequencing of ITS1/5.8S rRNA/ITS2 and Fe-hydrogenase as tools for genetic classification of potentially pathogenic isolates of Trichomonas gallinae.

作者信息

Sansano-Maestre José, Martínez-Herrero María Del Carmen, Garijo-Toledo María Magdalena, Gómez-Muñoz María Teresa

机构信息

Universidad Católica de Valencia, Facultad de Veterinaria y Ciencias Experimentales, Departamento de Producción Animal y Salud Pública, Calle Guillem de Castro, 94, 46003, Valencia, Spain.

Universidad CEU Cardenal Herrera, Instituto de Ciencias Biomédicas, Facultad de Veterinaria, Departamento de Producción y Sanidad Animal, Salud Pública Veterinaria y Ciencia y Tecnología de los Alimentos, Calle Tirant Lo Blanc, 7, 46115 Alfara del Patriarca, Valencia, Spain.

出版信息

Res Vet Sci. 2016 Aug;107:182-189. doi: 10.1016/j.rvsc.2016.05.016. Epub 2016 May 31.

Abstract

Trichomonas gallinae is a worldwide parasite that causes oropharyngeal avian trichomonosis. During eight years, 60 axenic isolates were obtained from different bird species and characterized by three molecular methods: RAPD analysis and PCR-sequencing of ITS1/5.8S rRNA/ITS2 fragment and Fe-hydrogenase gene. We have found two genotypes of ITS1/5.8S rRNA/ITS2 widely distributed among bird populations, a new variant and also two sequences with mixed pattern. Genotype ITS-OBT-Tg-1 was associated with the presence of gross lesions in birds. We have found eight genotypes of the Fe-hydrogenase (A1, A2, C2, C2.1, C4, C5, C6 and C7), three of them are new reports (C5, C6 and C7), and also three sequences with mixed pattern. Subtype A1 of the Fe-hydrogenase was also related with the presence of lesions. RAPD analyses included most of the strains isolated from animals with lesions in one of the sub-clusters. Potentially pathogenic isolates of T. gallinae obtained in this study fulfill the following criteria with one exception: isolated from lesions+ITS-OBT-Tg-1 genotype+FeHyd A1+RAPD sub-cluster I2.

摘要

鸽毛滴虫是一种可导致口咽型禽毛滴虫病的全球寄生虫。在八年时间里,从不同鸟类物种中获得了60株无菌分离株,并通过三种分子方法进行了特征分析:RAPD分析以及对ITS1/5.8S rRNA/ITS2片段和铁氢化酶基因的PCR测序。我们发现ITS1/5.8S rRNA/ITS2的两种基因型在鸟类群体中广泛分布,一种新变体以及两个具有混合模式的序列。基因型ITS-OBT-Tg-1与鸟类中明显病变的存在相关。我们发现了铁氢化酶的八种基因型(A1、A2、C2、C2.1、C4、C5、C6和C7),其中三种是新报道(C5、C6和C7),还有三个具有混合模式的序列。铁氢化酶的A1亚型也与病变的存在有关。RAPD分析包括从一个亚群中患有病变的动物分离出的大多数菌株。本研究中获得的潜在致病性鸽毛滴虫分离株符合以下标准,但有一个例外:从病变中分离+ITS-OBT-Tg-1基因型+FeHyd A1+RAPD亚群I2。

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