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Please use this identifier to cite or link to this item: http://hdl.handle.net/10400.18/202

Título: Development of a PCR-RFLP marker to genetically distinguish Prosorhynchus crucibulum and Prosorhynchus aculeatus
Autor: Francisco, Claire Juliana
Almeida, André
Castro, António Manuel
Santos, Maria João
Palavras-chave: Prosorhynchus crucibulum
Prosorhynchus aculeatus
18S rDNA region
Molecular identification
Infecções Sistémicas e Zoonoses
Issue Date: Mar-2010
Editora: Elsevier
Citação: Parasitol Int. 2010 Mar;59(1):40-3. Epub 2009 Oct 8
Resumo: The cercariae stages of Prosorhynchus crucibulum and Prosorhynchus aculeatus are morphologically indistinguishable. However, the differentiation of these two species is crucial to understand the transmission dynamics between these primary hosts (mussels) and the secondary hosts (fish). In this way, the objective of this study is to develop an accurate molecular identification tool to differentiate the cercariae stage of P. crucibulum and P. aculeatus. We targeted the 18S nuclear ribosomal DNA region by PCR amplification and sequenced this amplicon. By generating these sequences, we developed a RFLP tool with the use of the enzymes HincII and FokI that produced different restriction profiles between P. crucibulum and P. aculeatus. Each enzyme generated different-sized fragments specific to the species examined and no cross-reaction between the species was detected in their restriction pattern. By sequencing, no intraspecific-polymorphism was detected since there is 100% homology among P. aculeatus or P. crucibulum. These results indicate that PCR-linked restriction analysis of the 18S rDNA region provided us with rapid and reliable molecular tools for distinction of the cercariae of these species. The sequences generated were deposited in GenBank accession numbers for P. crucibulum cercariae (FJ463407, FJ463408 and FJ463409) and adult worm (FJ429096, FJ429097), and for P. aculeatus adult (FJ429094 and FJ429095).
Arbitragem científica: yes
URI: http://hdl.handle.net/10400.18/202
ISSN: 1383-5769
Versão do Editor: http://www.sciencedirect.com/science/article/pii/S1383576909001147
Appears in Collections:DDI - Artigos em revistas internacionais

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