title: Comparative assessment of ELISAs using recombinant saposin-like protein 2 and recombinant cathepsin L-1 from Fasciola hepatica for the serodiagnosis of human Fasciolosis.
abstract: Two recombinant Fasciola hepatica antigens, saposin-like protein-2 (recSAP2) and cathepsin L-1 (recCL1), were assessed individually and in combination in enzyme-linked immunosorbent assays (ELISA) for the specific serodiagnosis of human fasciolosis in areas of low endemicity as encountered in Central Europe. Antibody detection was conducted using ProteinA/ProteinG (PAG) conjugated to alkaline phosphatase. Test characteristics as well as agreement with results from an ELISA using excretory-secretory products (FhES) from adult stage liver flukes was assessed by receiver operator characteristic (ROC) analysis, specificity, sensitivity, Youdens J and overall accuracy. Cross-reactivity was assessed using three different groups of serum samples from healthy individuals (n=20), patients with other parasitic infections (n=87) and patients with malignancies (n=121). The best combined diagnostic results for recombinant antigens were obtained using the recSAP2-ELISA (87% sensitivity, 99% specificity and 97% overall accuracy) employing the threshold (cut-off) to discriminate between positive and negative reactions that maximized Youdens J. The findings showed that recSAP2-ELISA can be used for the routine serodiagnosis of chronic fasciolosis in clinical laboratories; the use of the PAG-conjugate offers the opportunity to employ, for example, rabbit hyperimmune serum for the standardization of positive controls.
authors.affiliation: Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, Bern, Switzerland.
authors.forename: Bruno
authors.initials: B
authors.lastname: Gottstein
authors.affiliation: Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, Bern, Switzerland.
authors.forename: Marianne
authors.initials: M
authors.lastname: Schneeberger
authors.affiliation: Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, Bern, Switzerland.
authors.forename: Ghalia
authors.initials: G
authors.lastname: Boubaker
authors.affiliation: Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, Bern, Switzerland.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Merkle
authors.affiliation: Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, Bern, Switzerland.
authors.forename: Cristina
authors.initials: C
authors.lastname: Huber
authors.affiliation: Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, Bern, Switzerland.
authors.forename: Markus
authors.initials: M
authors.lastname: Spiliotis
authors.affiliation: Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, Bern, Switzerland.
authors.forename: Norbert
authors.initials: N
authors.lastname: Müller
authors.affiliation: Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain.
authors.forename: Teresa
authors.initials: T
authors.lastname: Garate
authors.affiliation: Veterinary Public Health Institute, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
authors.forename: Marcus G
authors.initials: MG
authors.lastname: Doherr
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000947:Antigens, Helminth; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005060:Europe; D005210:Fasciola hepatica / Q000276:immunology*; D005211:Fascioliasis / Q000175:diagnosis*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods; D055815:Young Adult
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D011994:Recombinant Proteins