title: A novel monoclonal antibody-based immunoenzymatic assay for epidemiological surveillance of the vector snails of Fasciola hepatica (Trematoda: Digenea).
abstract: Fasciolosis is a globally distributed snail-borne disease which requires economic consideration due to its enormous impact on veterinary medicine. During recent decades, this parasitosis has also shown increasing prevalence in human populations worldwide. The dissemination and successful transmission of fasciolosis ultimately depends on the existence of susceptible snails that act as intermediate hosts. Therefore, to accomplish effective control of this disease, surveillance and detection of the infected intermediate host would be essential. The screening of trematodes within snails using classical parasitological examination of the larvae can be unreliable (sensitivity and specificity vary depending on the time of infection and the experience of the observer) and relatively costly when using molecular biological methods during large-scale monitoring. Here we propose a novel monoclonal antibody-based immunoenzymatic assay to detect ongoing Fasciola hepatica infection in lymnaeid snails. Anti-F. hepatica rediae mouse monoclonal antibodies were generated and used to develop a double monoclonal antibody-based ELISA for parasite detection. Fasciola hepatica-infected and uninfected laboratory-reared Galba cubensis and Pseudosuccinea columella were used for assessment of the developed ELISA. Experimentally infected snails were dissected and examined for parasite larvae as the "gold standard" method. Sensitivity results were 100% for both snail species, while specificity was 98% for G. cubensis and 100% for P. columella. No cross-reactivity was detected in lymnaeids infected with Trichobilharzia sp. or Cotylophoron sp. The ELISA enabled detection of the infection from day 8 p.i. in G. cubensis while in P. columella it was noted as early as day 4. To our knowledge no previous immunoassays have been reported to detect helminth-infected snails and the developed sandwich ELISA method is therefore suggested for infection status validation in natural populations of lymnaeid snails.
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6½, La Lisa, AP 601 La Habana, Cuba. Electronic address: annia@ipk.sld.cu.
authors.forename: Annia
authors.initials: A
authors.lastname: Alba
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6½, La Lisa, AP 601 La Habana, Cuba.
authors.forename: Hilda M
authors.initials: HM
authors.lastname: Hernández
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6½, La Lisa, AP 601 La Habana, Cuba.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Marcet
authors.affiliation: Laboratorio de Malacología, Departamento de Control de Vectores, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6½, 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 6½, La Lisa, AP 601 La Habana, Cuba.
authors.forename: Mabel
authors.initials: M
authors.lastname: Figueredo
authors.affiliation: Laboratorio de Malacología, Departamento de Control de Vectores, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6½, AP 601 La Habana, Cuba.
authors.forename: Jorge
authors.initials: J
authors.lastname: Sánchez
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Vicepresidencia de Investigaciones, Instituto Finlay, Ave 198, No. 19805, AP 11600 La Habana, Cuba.
authors.forename: Oscar
authors.initials: O
authors.lastname: Otero
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Ave Novia del Mediodía km 6½, La Lisa, AP 601 La Habana, Cuba.
authors.forename: Jorge
authors.initials: J
authors.lastname: Sarracent
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth / Q000032:analysis*; D004199:Disease Vectors; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D062665:Epidemiological Monitoring; D005210:Fasciola hepatica / Q000276:immunology / Q000302:isolation & purification*; D005260:Female; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity; D012908:Snails / Q000469:parasitology*
chemical_list: D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth
keywords: ELISA; Epidemiology; Fasciolosis; Lymnaeidae; Snail-borne disease control