title: A multiplex PCR for the detection of Fasciola hepatica in the intermediate snail host Galba cubensis.
abstract: Fasciolosis is a snail-borne trematode infection that has re-emerged as a human disease, and is considered a significant problem for veterinary medicine worldwide. The evaluation of the transmission risk of fasciolosis as well as the efficacy of the strategies for its control could be carried out through epidemiological surveillance of the snails that act as intermediate hosts of the parasites. The present study aimed to develop the first multiplex PCR to detect Fasciola hepatica in Galba cubensis, an important intermediate host of the parasite in the Americas and especially in the Caribbean basin. The multiplex PCR was optimized for the amplification of a 340 bp fragment of the second internal transcribed spacer (ITS-2) of F. hepatica rDNA, while another set of primers was designed and used to amplify a conserved segment of the nuclear 18S rDNA of the snail (451 bp), as an internal control of the reaction. The assay was able to detect up to 100 pg of the parasite even at high concentrations of snail DNA, an analytical sensitivity that allows the detection of less than a single miracidium, which is the minimal biological infestation unit. A controlled laboratory-reared G. cubensis - F. hepatica system was used for the evaluation of the developed multiplex PCR, and 100% sensitivity and specificity was achieved. This assay constitutes a novel, useful and suitable technique for the survey of fasciolosis transmission through one of the main intermediate hosts in the Western hemisphere.
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba.
authors.forename: Annia
authors.initials: A
authors.lastname: Alba
authors.affiliation: Laboratorio de Malacología, Departamento de Control de Vectores, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba.
authors.forename: Antonio A
authors.initials: AA
authors.lastname: Vázquez
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba.
authors.forename: Hilda
authors.initials: H
authors.lastname: Hernández
authors.affiliation: Laboratorio de Malacología, Departamento de Control de Vectores, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba.
authors.forename: Jorge
authors.initials: J
authors.lastname: Sánchez
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Marcet
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba.
authors.forename: Mabel
authors.initials: M
authors.lastname: Figueredo
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba.
authors.forename: Jorge
authors.initials: J
authors.lastname: Sarracent
authors.affiliation: Laboratorio de Biología Molecular, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6 ½, AP 601, La Habana, Cuba. Electronic address: fraga@ipk.sld.cu.
authors.forename: Jorge
authors.initials: J
authors.lastname: Fraga
mesh_terms: D000818:Animals; D018509:DNA, Helminth / Q000235:genetics; D005210:Fasciola hepatica / Q000302:isolation & purification / Q000502:physiology*; D006790:Host-Parasite Interactions; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012908:Snails / Q000469:parasitology*
chemical_list: D018509:DNA, Helminth; D012337:RNA, Ribosomal, 18S
keywords: Diagnosis; Epidemiology; Fasciola hepatica; Galba cubensis; Multiplex PCR