title: An ELISA using recombinant TmHSP70 for the diagnosis of Taenia multiceps infections in goats.
abstract: Infections with the tapeworm Taenia multiceps are problematic for ruminant farming worldwide. Here we develop a novel and rapid method for serodiagnosis of T. multiceps infections via an indirect ELISA (iELISA) that uses a heat shock protein, namely, TmHSP70. We extracted the total RNA of T. multiceps from the protoscoleces of cysts dissected from the brains of infected goats. Subsequently, we successfully amplified, cloned and expressed the TmHSP70 gene in Escherichia coli BL21 (DE3). Western blot analysis showed that the recombinant protein (∼34 kDa molecular weight) was recognized by the coenurosis positive serum. Given these initial, robust immunogenic properties for recombinant TmHSP protein, we assessed the ELISA-based serodiagnostic potential of this gene. The indirect ELISA was then optimized to 2.70 μg/well dilution for antigen and 1:80 dilution for serum,while the cut-off value is 0.446. We report that our novel TmHSP ELISA detected T. multiceps sera with a sensitivity of 1:10240 and a specificity of 83.3% (5/6). In a preliminary application, this assay correctly confirmed T. multiceps infection in 30 infected goats, consistent with the clinical examination. This study has revealed that our novel iELISA, which uses the rTmHSP protein, provides a rapid test for diagnosing coenurosis.
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Huaming
authors.initials: H
authors.lastname: Nie
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Xiaobin
authors.initials: X
authors.lastname: Gu
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Weimin
authors.initials: W
authors.lastname: Lai
authors.affiliation: Department of Chemistry, College of Science, Sichuan Agricultural University, Ya'an 625014, China.
authors.forename: Xuerong
authors.initials: X
authors.lastname: Peng
authors.affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China. Electronic address: guangyou1963@aliyun.com.
authors.forename: Guangyou
authors.initials: G
authors.lastname: Yang
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth; D003001:Cloning, Molecular; D003551:Cysticercosis / Q000175:diagnosis / Q000662:veterinary*; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D015511:Goat Diseases / Q000175:diagnosis / Q000469:parasitology*; D006041:Goats; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology*; D015801:Helminth Proteins / Q000276:immunology*; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013621:Taenia / Q000145:classification*
chemical_list: D000909:Antibodies, Helminth; D018076:DNA, Complementary; D018840:HSP70 Heat-Shock Proteins; D015801:Helminth Proteins; D011994:Recombinant Proteins
keywords: Clone; Expression; Immunodiagnosis; Indirect ELISA; Taenia multiceps; Tapeworm; TmHSP 70