Departamento de Zootecnia, Universidade Federal de Minas Gerais, Escola de Veterinária, Av. Antônio Carlos 6627, Pampulha, Belo Horizonte, MG, 31270901, Brazil.
Laboratório de Biologia Celular e Tecidual, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Av. Alberto Lamego, 2000, Parque Califórnia, Campos dos Goytacazes, RJ, 28013602, Brazil.
Parasitol Res. 2020 Feb;119(2):529-543. doi: 10.1007/s00436-019-06569-3. Epub 2019 Dec 13.
Standard diagnostic methods currently in use for the identification of helminth infections in ruminants are based on the morphological analysis of immature and adult stages of parasites. This paper describes a method for the semiquantitative identification of nematodes, mainly Trichostrongyloidea, at species-level resolution. The method is based on amplification and fragment analysis followed by minisequencing of the ITS-2 region (internal transcribed spacer 2) of the ribosomal DNA of parasite eggs or larvae. This method allows for the identification of seven genera (Chabertia, Cooperia, Haemonchus, Oesophagostomum, Ostertagia, Teladorsagia, and Trichostrongylus) and 12 species (Chabertia ovina, Cooperia curticei, Cooperia punctata, Cooperia oncophora/Cooperia surnabada, Haemonchus contortus, Haemonchus placei, Haemonchus longistipes, Oesophagostomum asperum, Oesophagostomum radiatum, Ostertagia ostertagi, Trichostrongylus axei, and Trichostrongylus colubriformis) of infectious nematodes of domestic ruminants. The concordance between the morphological and molecular analyses in the detection of genera ranged from 0.84 to 0.99, suggesting the proposed detection method is specific, semiquantitative, less laborious, and highly cost-efficient.
目前用于鉴定反刍动物寄生虫感染的标准诊断方法基于对寄生虫未成熟和成虫阶段的形态分析。本文描述了一种用于半定量鉴定线虫（主要是 Trichostrongyloidea）到种水平的方法。该方法基于寄生虫卵或幼虫核糖体 DNA 的 ITS-2 区域（内部转录间隔区 2）的扩增和片段分析，随后进行 minisequencing。该方法可鉴定七个属（Chabertia、Cooperia、Haemonchus、Oesophagostomum、Ostertagia、Teladorsagia 和 Trichostrongylus）和 12 个种（Chabertia ovina、Cooperia curticei、Cooperia punctata、Cooperia oncophora/Cooperia surnabada、Haemonchus contortus、Haemonchus placei、Haemonchus longistipes、Oesophagostomum asperum、Oesophagostomum radiatum、Ostertagia ostertagi、Trichostrongylus axei 和 Trichostrongylus colubriformis）的感染性反刍动物线虫。在检测属时，形态学和分子分析之间的一致性在 0.84 到 0.99 之间，表明所提出的检测方法具有特异性、半定量、劳动强度低和高成本效益。
