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Isolation of two lipoprotein antigens from the metacestodes of Taenia hydatigena (Pallas, 1766) and Taenia multiceps (Leske, 1780) and their evaluation in sero-diagnosis.

Two identical host contaminant-free lipoprotein antigens were isolated from the metacestodes of T. hydatigena and T. multiceps, of which antigen 1 was more reactive than antigen 2. These antigens appear to be identical with antigens B and A, respectively, and antigen 2 with arc 5 of Echinococcus granulosus described by other workers. Attempts to use these antigens in serodiagnosis showed that only in the indirect haemagglutination (IHA) technique was there any increased sensitivity and specificity compared with the unpurified cyst fluid antigens. No correlation was found between the size of the infecting dose, the number of cysts found at necropsy, the IHA titres, or number of precipitation lines seen in immunodiffusion tests.

Animals↗

PCR tools for the differential diagnosis of Taenia saginata and Taenia solium taeniasis/cysticercosis from different geographical locations.

The potential value of PCRs in the species-specific diagnosis of have been investigated, using samples of T. saginata and T. solium from different geographical areas. The PCRs examining inter-species differences were based on the sequence of the HDP2 DNA fragment, specific for T. saginata/T. solium, and the sequence of the rDNA internal transcribed spacer 1 and spacer 2 (ITS-1 and ITS-2). This PCR analysis of DNA isolates confirmed morphologic diagnosis and allowed the speciation of samples too small or fragmented for morphologic identification, with clear and consistent inter-species differences between T. saginata (twenty-two) and T. solium (three) geographical isolates. Possible intra-species genomic variability, within these species, was similarly studied through analysis of PCR amplification products (PCR-RFLP) and only encountered one exceptional T. saginata isolate from Kenya, which yielded a unique PCR-RFLP pattern, different from T. saginata DNA of Mexican (one sample) and Spanish (seven samples) origin.

Animals↗

Taenia crassiceps metacestode vesicular fluid antigens shared with the Taenia solium larval stage and reactive with serum antibodies from patients with neurocysticercosis.

After removal of host (mouse) serum proteins (albumin, transferrin, IgG and another five unidentified proteins) by immunoaffinity chromatography, the vesicular fluid of T. crassiceps metacestodes (TcVF) was immunoelectrophoretically examined for antigens recognized by rabbit antiserum to aqueous crude extract of T. solium cysticerci. Based on a precipitate pattern developed in electroimmunodiffusion, nine cross-reactive antigens could be demonstrated. In the ELISA, TcVF was shown to be a potent antigen for the demonstration of IgG antibodies in the sera of Mexican patients (n = 14) with confirmed neurocysticercosis (mean E490 values +/- SD: 0.39 +/- 0.38) although it was less sensitive when compared to T. solium VF (0.95 +/- 0.54). Sensitivity was much higher using cross-reactive TcVF antigens selected by immunoaffinity chromatography with rabbit IgG antibodies to larval T. solium crude extract (0.87 +/- 0.57). SDS-PAGE fluorograph of cross-reactive, radioiodinated TcVF protein antigens and selected by antibodies of individual neurocysticercosis sera (n = 13), exhibited six to nine bands depending on the serum tested. Altogether ten 125I-labelled proteins (Mr range from about 20,000 to 200,000) were recognized by neurocysticerosis antibodies. Four proteins (Mr about 22,000, 25,000, 32,000 and 45,000) were detected by all sera.

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Production of monoclonal antibodies anti-Taenia crassiceps cysticerci with cross-reactivity with Taenia solium antigens.

We describe the production of the potential monoclonal antibodies (MoAbs) using BALB/c mice immunized with vesicular fluid (VF)-Tcra (T. crassiceps) antigen. Immune sera presented anti-VF-Tcra (<20kD) IgG and IgM antibodies with cross-reactivity with T. solium (Tso) antigen (8-12, 14, and 18 kD). After cell fusion, we selected 33 anti-Tcra and anti-Tso reactive IgM-clones and 53 anti-Tcra specific IgG-clones, 5 of them also recognizing Tso antigens. Two clones identified the 8-14 and 18kD peptides of VF-Tcra.

Animals↗