Alda Pilar, Lounnas Manon, Vázquez Antonio Alejandro, Ayaqui Rolando, Calvopiña Manuel, Celi-Erazo Maritza, Dillon Robert T, Jarne Philippe, Loker Eric S, Muñiz Pareja Flavia Caroll, Muzzio-Aroca Jenny, Nárvaez Alberto Orlando, Noya Oscar, Robles Luiggi Martini, Rodríguez-Hidalgo Richar, Uribe Nelson, David Patrice, Pointier Jean-Pierre, Hurtrez-Boussès Sylvie
Laboratorio de Zoología de Invertebrados I, Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur, San Juan No. 670, B8000ICN, Bahía Blanca, Buenos Aires, Argentina; MIVEGEC, University of Montpellier, CNRS, IRD, Montpellier, France.
MIVEGEC, University of Montpellier, CNRS, IRD, Montpellier, France.
Vet Parasitol. 2018 Feb 15;251:101-105. doi: 10.1016/j.vetpar.2018.01.006. Epub 2018 Jan 8.
A molecular tool described here allows in one step for specific discrimination among three cryptic freshwater snail species (genus Galba) involved in fasciolosis transmission, a worldwide infectious disease of humans and livestock. The multiplex PCR approach taken targets for each species a distinctive, known microsatellite locus which is amplified using specific primers designed to generate an amplicon of a distinctive size that can be readily separated from the amplicons of the other two species on an agarose gel. In this way, the three Galba species (G. cubensis, G. schirazensis, and G. truncatula) can be differentiated from one another, including even if DNA from all three were present in the same reaction. The accuracy of this new molecular tool was tested and validated by comparing multiplex PCR results with species identification based on sequences at mitochondrial and nuclear markers. This new method is accurate, inexpensive, simple, rapid, and can be adapted to handle large sample sizes. It will be helpful for monitoring invasion of Galba species and for developing strategies to limit the snail species involved in the emergence or re-emergence of fasciolosis.
本文所述的一种分子工具能够一步实现对三种隐秘淡水螺（Galba属）的特异性区分，这三种淡水螺参与了肝片吸虫病的传播，而肝片吸虫病是一种影响人类和牲畜的全球性传染病。所采用的多重PCR方法针对每个物种的一个独特的、已知的微卫星位点，使用特定引物进行扩增，这些引物设计用于产生具有独特大小的扩增子，该扩增子能够在琼脂糖凝胶上轻松地与其他两个物种的扩增子分离。通过这种方式，三种Galba属物种（古巴Galba、设拉子Galba和截形Galba）能够彼此区分开来，即使在同一反应中存在来自所有三个物种的DNA也能做到。通过将多重PCR结果与基于线粒体和核标记序列的物种鉴定结果进行比较，对这种新分子工具的准确性进行了测试和验证。这种新方法准确、廉价、简单、快速，并且能够适用于处理大量样本。它将有助于监测Galba属物种的入侵情况，并有助于制定策略来限制参与肝片吸虫病出现或再次出现的螺类物种。
