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Assessment of the exposure to Echinococcus multilocularis associated with carnivore faeces using real-time quantitative PCR and flotation technique assays.
International Journal for Parasitology ( IF 4 ) Pub Date : 2020-09-04 , DOI: 10.1016/j.ijpara.2020.07.008
Abdou Malik Da Silva 1 , Sandra Courquet 2 , Francis Raoul 1 , Dominique Rieffel 1 , Patrick Giraudoux 1 , Laurence Millon 2 , Jenny Knapp 2
Affiliation  

The eggs of Echinococcus multilocularis, the infectious stage, are spread into the environment through wild and domestic carnivore faeces. The spatial location of the faeces containing infective E. multilocularis eggs is a key parameter for studying areas of exposure and understanding the transmission processes to the intermediate hosts and humans. Echinococcus multilocularis faecal prevalence is often assessed by detecting E. multilocularis DNA, not necessarily eggs. This work aimed to determine the percentage of faeces containing E. multilocularis eggs in a rural town and its surroundings and whether this level of precision is relevant in assessing exposure to E. multilocularis. For this purpose, we developed a combined molecular and microscopic approach to investigate the E. multilocularis exposure of potential hosts in the environment from field-collected carnivore faeces. Carnivore defecation patterns were then spatialized to study the spatial distribution of E. multilocularis. Faeces were screened for E. multilocularis DNA using a specific real-time quantitative PCR (qPCR). Echinococcus multilocularis eggs were morphologically identified from E. multilocularis-specific qPCR-positive faeces after sucrose flotation and individually confirmed through specific PCR and sequencing. The spatial distribution of E. multilocularis was studied using Kulldorff statistics. Echinococcus multilocularis eggs were identified mostly in fox faeces positive for E. multilocularis DNA by qPCR (n = 27/70) and only from 1 of 15 copro-samples from dogs and 1 of 5 from cats. The faecal prevalence of E. multilocularis DNA and eggs was overdispersed, with the same geographical patterns. These data suggest that E. multilocularis DNA and/or egg detection in carnivore faeces, mainly that of foxes, is appropriate in ecological studies of E. multilocularis transmission.



中文翻译:

使用实时定量 PCR 和浮选技术分析评估与食肉动物粪便相关的多房棘球绦虫的暴露情况。

房棘球绦虫的卵通过野生和家养食肉动物的粪便传播到环境中。含有感染性多房大肠杆菌卵的粪便的空间位置是研究暴露区域和了解中间宿主和人类传播过程的关键参数。多房棘球绦虫粪便流行率通常通过检测多棘球蚴DNA(不一定是鸡蛋)来评估。这项工作旨在确定农村城镇及其周围环境中含有多房大肠杆菌卵的粪便百分比,以及该精度水平是否与评估多房大肠杆菌暴露相关. 为此,我们开发了一种分子和微观相结合的方法,以研究来自现场收集的食肉动物粪便的环境中潜在宿主的多房大肠杆菌暴露。然后将食肉动物的排便模式空间化以研究多球蚴的空间分布。使用特定的实时定量 PCR (qPCR)筛选粪便中的多房大肠杆菌DNA。多房棘球蚴卵在蔗糖浮选后从多棘球蚴特异性 qPCR 阳性粪便中进行形态学鉴定,并通过特异性 PCR 和测序进行单独确认。多球蚧的空间分布使用 Kulldorff 统计进行研究。棘球绦虫卵大多在狐被鉴定为阳性的粪便E.多房通过qPCR DNA(Ñ  =70分之27),只有1从犬5的15的Copro从样本和1从猫。E. multilocularis DNA 和鸡蛋的粪便流行过度分散,具有相同的地理模式。这些数据表明,肉食动物粪便(主要是狐狸的粪便)中的多房大肠杆菌DNA 和/或卵检测适用于多房大肠杆菌传播的生态学研究。

更新日期:2020-09-04
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