Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR.
From complete mitochondrial DNA sequence of Fasciola hepatica available in Genbank, specific primers were designed for a conserved and repetitive region of this trematode. A pair of primers was used for diagnosis of infected Lymnaea columella by F. hepatica during the pre-patent period simultaneously with another pair of primers which amplified the internal transcribed spacer (ITS) region of rDNA from L. columella in a single Multiplex-PCR. The amplification generated a ladder band profile specific for F. hepatica. This profile was observed in positive molluscs at different times of infection, including adult worms from the trematode. The Multiplex-PCR technique showed to be a fast and safe tool for fascioliasis diagnosis, enabling the detection of F. hepatica miracidia in L. columella during the pre-patent period and identification of transmission areas.
Centro de Pesquisas René Rachou-Fiocruz, Av. Augusto de Lima 1715, 30190-002 Belo Horizonte, MG, Brazil.
Kelly Grace
KG
Magalhães
Liana Konovaloff Jannotti
LK
Passos
Omar dos Santos
Odos S
Carvalho
D000818:Animals; D018509:DNA, Helminth / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005210:Fasciola hepatica / Q000235:genetics / Q000302:isolation & purification*; D008195:Lymnaea / Q000235:genetics / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*
D018509:DNA, Helminth; D004272:DNA, Mitochondrial; D021903:DNA, Ribosomal Spacer