title: Evaluation of a novel Dot-ELISA assay utilizing a recombinant protein for the effective diagnosis of Taenia pisiformis larval infections.
abstract: Cysticercosis, caused by the larvae of Taenia pisiformis, is a common disease in domestic breeds of the rabbit Oryctolagus cuniculus that results in economic losses. At present, there is no convenient and effective method for the rapid detection of T. pisiformis larvae. Here, we developed and tested the efficacy of a Dot-ELISA assay for the diagnosis of T. pisiformis larval infections in rabbits, based on the expression of the recombinant fusion protein (rTp1) from the Tp1 gene. Rapid amplification of cDNA ends (RACE) was used to amplify the 3' ends of the Tp1 gene, based on the unigene similar to Ts1 gene (EU009656.1) which comes from transcriptome sequencing of T. pisiformis. The Tp1 gene was successfully amplified, cloned and expressed in BL21 (DE3). Western blot analysis revealed that the recombinant Tp1 protein is specifically recognized by rabbit T. pisiformis cysticercosis antisera. This purified recombinant fusion protein, rTp1, was probed by Dot-ELISA with sera from rabbits infected with T. pisiformis larvae and with other parasitic infections. Results showed that this Dot-ELISA assay had both high sensitivity (92.9-97.6%) and specificity (95.2-98.4%) to detect T. pisiformis larval infections. We also found very low levels of cross-reaction with other parasitic infections. This study has revealed that our novel Dot-ELISA assay utilizing the recombinant fusion protein, rTp1, has a strong potential for the effective diagnosis of T. pisiformis infections in rabbits.
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: Deying
authors.initials: D
authors.lastname: Yang
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: College of Life and Basic 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: D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000947:Antigens, Helminth / Q000276:immunology*; D003551:Cysticercosis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D003552:Cysticercus / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D007814:Larva; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins; D012680:Sensitivity and Specificity; D016415:Sequence Alignment / Q000662:veterinary; D017422:Sequence Analysis, DNA; D013621:Taenia / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D013622:Taeniasis / Q000276:immunology
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins
keywords: Clone; Dot-ELISA; Expression; Immunodiagnosis; Taenia pisiformis; Tp1