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用于诊断马感染叶状裸头绦虫(绦虫纲,圆叶目)的粪便学、血清学和分子方法的比较

A comparison of coprological, serological and molecular methods for the diagnosis of horse infection with Anoplocephala perfoliata (Cestoda, Cyclophyllidea).

作者信息

Traversa Donato, Fichi Gianluca, Campigli Michela, Rondolotti Anna, Iorio Raffaella, Proudman Christopher J, Pellegrini Duccio, Perrucci Stefania

机构信息

Department of Comparative Biomedical Sciences, Faculty of Veterinary Medicine, University of Teramo, Italy.

出版信息

Vet Parasitol. 2008 Apr 15;152(3-4):271-7. doi: 10.1016/j.vetpar.2007.12.032. Epub 2008 Jan 8.

Abstract

Anoplocephala perfoliata (Cestoda, Cyclophyllidea), the commonest intestinal tapeworm of horses, can cause colic, intussusceptions, ileal impactions and intestinal perforations. Common diagnostic techniques for A. perfoliata infection, i.e. coprology and serology, show inherent limitations in terms of sensitivity and specificity and new approaches are thus required. Hence, the present study compared the reliability of coprological, serological (i.e. ELISA) and molecular (i.e. nested PCR) methods in detecting A. perfoliata infection in naturally infected horses and in horses treated with a combination of ivermectin and praziquantel. Of 42 horses subjected to coprological examination, 16 and 26 resulted negative and positive, respectively for the presence of A. perfoliata eggs at the coprological examination. The 26 coprologically positive animals were also positive by nested PCR. Fifteen out of the 16 horses coprologically negative were negative at the molecular assay, while one yielded a PCR product detectable on an agarose gel. Eighteen out of 26 positive horses were treated with a combination of ivermectin 18.7 mg/g and praziquantel 140.3mg/g and resulted subsequently negative by coprology and nested PCR performed 2 weeks after treatment. All infected and untreated animals had a high ELISA test optical density indicating high infection intensity and associated risk of colic. However, high optical density values were also obtained in four horses post-treatment and in three horses that were negative on molecular and coprological analysis. The results of the present work indicate that the nested PCR assay represents a valid method for the specific molecular detection of A. perfoliata in faecal samples collected from naturally infected horses and may have advantages over coprological and serological approaches for diagnosing A. perfoliata infection.

摘要

叶状裸头绦虫(Anoplocephala perfoliata,绦虫纲,圆叶目)是马最常见的肠道绦虫,可引起绞痛、肠套叠、回肠阻塞和肠穿孔。叶状裸头绦虫感染的常用诊断技术,即粪便学和血清学方法,在敏感性和特异性方面存在固有局限性,因此需要新的方法。因此,本研究比较了粪便学、血清学(即酶联免疫吸附测定)和分子学(即巢式聚合酶链反应)方法在检测自然感染马和接受伊维菌素与吡喹酮联合治疗的马的叶状裸头绦虫感染方面的可靠性。在接受粪便学检查的42匹马中,粪便学检查时,分别有16匹和26匹的叶状裸头绦虫虫卵呈阴性和阳性。26匹粪便学检查呈阳性的动物通过巢式聚合酶链反应也呈阳性。16匹粪便学检查呈阴性的马中有15匹在分子检测中呈阴性,而有1匹产生了在琼脂糖凝胶上可检测到的聚合酶链反应产物。26匹阳性马中有18匹接受了18.7mg/g伊维菌素和140.3mg/g吡喹酮的联合治疗,治疗2周后进行粪便学和巢式聚合酶链反应检测,结果随后呈阴性。所有感染且未治疗的动物酶联免疫吸附测定的光密度都很高,表明感染强度高且有发生绞痛的相关风险。然而,在4匹治疗后的马和3匹分子和粪便学分析呈阴性的马中也获得了高光密度值。本研究结果表明,巢式聚合酶链反应检测法是从自然感染马采集的粪便样本中特异性分子检测叶状裸头绦虫的有效方法,在诊断叶状裸头绦虫感染方面可能比粪便学和血清学方法更具优势。

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