title: Validation of a recombinant protein indirect ELISA for the detection of specific antibodies against Theileria uilenbergi and Theileria luwenshuni in small ruminants.
abstract: An enzyme-linked immunosorbent assay (ELISA) based on a recombinant Theileria uilenbergi immunodominant protein (rTuIP) was validated for detection of antibodies in 188 positive and 198 negative reference serum samples, respectively. The cut-off value was determined at 32.7% with 95% and 90% accuracy levels by two-graphic receiver-operating characteristic (TG-ROC). The equal diagnostic sensitivity (Se) and specificity (Sp) were calculated to be 98.4%. Further validation of the repeatability with positive and negative reference samples indicated the reliable performance of the assay. Monitoring the antibody dynamics of sheep experimentally infected with Theileria luwenshuni showed the efficient detection of antibody response against the pathogen at the early infection stage and up until two months post infection. Application of this assay for detection of antibody in field sera from previous unknown Theileria endemic regions in Suizhou and Guiyang showed 17.8% and 11.6% seroprevalence, respectively, and presence of the pathogen was confirmed by identification of the 18S rRNA gene in the corresponding blood of the seropositive animals. These data support that the rTuIP ELISA could be a useful tool to study the epidemiology of theileriosis caused by T. uilenbergi and/or T. luwenshuni.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases MOA, Key Laboratory of Veterinary Parasitology of Gansu Province, LVRI, CAAS, Lanzhou, PR China.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases MOA, Key Laboratory of Veterinary Parasitology of Gansu Province, LVRI, CAAS, Lanzhou, PR China.
authors.forename: Youquan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Veterinary Research Institute, Khartoum, Sudan.
authors.forename: Dia Eldin A
authors.initials: DE
authors.lastname: Salih
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases MOA, Key Laboratory of Veterinary Parasitology of Gansu Province, LVRI, CAAS, Lanzhou, PR China.
authors.forename: Jianxun
authors.initials: J
authors.lastname: Luo
authors.affiliation: Forschungszentrum Borstel, Parkallee 22, 23845 Borstel, Germany.
authors.forename: Jabbar S
authors.initials: JS
authors.lastname: Ahmed
authors.affiliation: Forschungszentrum Borstel, Parkallee 22, 23845 Borstel, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Seitzer
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Grazing Animal Diseases MOA, Key Laboratory of Veterinary Parasitology of Gansu Province, LVRI, CAAS, Lanzhou, PR China. Electronic address: yinhong@caas.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Yin
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D002681:China / Q000453:epidemiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D015800:Protozoan Proteins / Q000276:immunology*; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012418:Ruminants; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies; D012756:Sheep; D012757:Sheep Diseases / Q000453:epidemiology* / Q000276:immunology / Q000469:parasitology; D016794:Theileria / Q000235:genetics / Q000276:immunology*; D013801:Theileriasis / Q000453:epidemiology* / Q000276:immunology / Q000469:parasitology
chemical_list: D000913:Antibodies, Protozoan; D015800:Protozoan Proteins; D011994:Recombinant Proteins
keywords: Theileria luwenshuni; Theileria uilenbergi; Validation; rTuIP ELISA