Amarante M R V, Santos M C, Bassetto C C, Amarante A F T
Departamento de Parasitologia,Instituto de Biociências,UNESP - Universidade Estadual Paulista,Botucatu,SP,Brazil.
J Helminthol. 2017 Nov;91(6):757-761. doi: 10.1017/S0022149X16000882. Epub 2017 Jan 10.
Haemonchus contortus and Haemonchus placei are among the major parasites of small ruminants and cattle. Although infection with these nematodes is host-specific, with H. placei predominating in cattle and H. contortus in sheep, cross-infections are observed in areas where both parasites are sympatric, and hybrid offspring can occur. Therefore, a fast and precise method is required for differentiating the parasites. Identification based on spicule morphometry is the most common technique for differentiating Haemonchus species. However, because these measurements overlap between species, morphological analysis is insufficient for differentiating between helminth species. In this work, we present a reliable, conventional polymerase chain reaction (PCR)-based method that uses two species-specific primer pairs to differentiate between H. contortus and H. placei specimens and their hybrids. Each primer pair produces one single and distinct amplification band for each species, which enables the detection of hybrid specimens. These primer pairs were validated by testing eight different populations of H. contortus, H. placei and hybrids.
捻转血矛线虫和柏氏血矛线虫是小型反刍动物和牛的主要寄生虫。尽管这些线虫感染具有宿主特异性，柏氏血矛线虫主要感染牛，捻转血矛线虫主要感染绵羊，但在两种寄生虫同域分布的地区会观察到交叉感染，并且可能出现杂交后代。因此，需要一种快速且精确的方法来区分这些寄生虫。基于交合刺形态测量的鉴定是区分血矛线虫种类最常用的技术。然而，由于这些测量值在不同物种之间存在重叠，形态学分析不足以区分蠕虫种类。在这项工作中，我们提出了一种可靠的、基于传统聚合酶链反应（PCR）的方法，该方法使用两对物种特异性引物来区分捻转血矛线虫和柏氏血矛线虫标本及其杂交种。每对引物对每个物种都产生一个单一且独特的扩增条带，从而能够检测杂交标本。通过测试捻转血矛线虫、柏氏血矛线虫和杂交种的八个不同种群对这些引物对进行了验证。
