RESUMO
We report the analysis by ELISA of class and subclass antibody response against a total soluble extract from T. spiralis adult stage (TSE-A) during a year after the infection in 17 symptomatic trichinellosis patients (SI) and five asymptomatic individual (AI) involved in an outbreak of trichinellosis occurred in the State of Mexico. Serum samples from 20 healthy individuals (HI) and 24 patients with other parasitosis were included as control. All SI showed a polyisotypic antibody response against the TSE-A, during the infection. Higher response of IgA, IgE, IgM were detected in SI during the acute phase of the infection, but only IgE remained at high levels all along the infection. None or a lower reactivity against TSE-A was observed in sera from AI and from HI. Some patients with trichuriosis and ascariosis showed a higher cross-reactivity, against TSE-A when IgG and their subclasses were analyzed.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Isotipos de Imunoglobulinas/sangue , Trichinella spiralis/imunologia , Triquinelose/imunologia , Adulto , Animais , Formação de Anticorpos , Surtos de Doenças , Ensaio de Imunoadsorção Enzimática , Seguimentos , Humanos , Imunoglobulina A/sangue , Imunoglobulina E/sangue , Imunoglobulina M/sangue , México/epidemiologia , Camundongos , Camundongos Endogâmicos BALB C , Valores de Referência , Fatores de Tempo , Triquinelose/sangue , Triquinelose/epidemiologiaRESUMO
The countercurrent immunoelectrophoresis (CIE) test was performed as a diagnostic tool for bovine babesiosis using a partially purified soluble antigen obtained from a saline extract of B. bovis-infected bovine erythrocytes by 60% ammonium sulphate precipitation (Bb-ag), and it was compared with the indirect fluorescent antibody test (IFA). Antibodies against B. bovis were detected in all 3 experimentally infected calves at the 8th post-infection day by IFA while antibodies to Bb-ag at the 10th postinfection day by CIE. These antibodies persisted until the end of the observation period (90th day post-infection). 43 serum samples from 116 bovines coming from a Babesia-enzootic area were positive (antibody titers > 1:30) by IFA test. 36 from the 43 (94%) IFA -positive bovine serum samples had antibodies to B. bovis soluble antigen (Bb-ag) detected by CIE. In contrast, the 20 serum samples from a Babesia-free area were all negative by both these methods. No cross-reactivity was observed with the soluble antigen obtained from normal red blood cells in the 3 experimentally infected calves nor in the bovines coming from a Babesia-enzootic area. CIE employing Bb-ag was as specific as IFA test but slightly less sensitive (100 vs. 94%) and detected antibodies to B.bovis early during the infection (8 vs. 10 days post-infection). Since CIE is easy to perform, does not require expensive reagents, material or equipment, and one can analyze up to 50 serum samples at once in a short time it is recommended as a useful test for the diagnosis of bovine babesiosis in developing countries.
Assuntos
Babesiose/diagnóstico , Doenças dos Bovinos/diagnóstico , Contraimunoeletroforese , Animais , Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Babesia/imunologia , Babesiose/sangue , Bovinos , Doenças dos Bovinos/sangue , Imunofluorescência , Estudos Longitudinais , Testes SorológicosRESUMO
Human antibody response to total soluble extract of Trichinella spiralis muscle larvae (TSE) was analyzed by Western blot. The most frequently recognized antigens had molecular weights of 96, 67, 63, 60, 55 and 47 kDa. An antigenic fraction containing two peptides with M.W. of 43, 47 kDa from the parasite (p43, 47 Ts L1) was isolated by elution from polyacrylamide gel slabs. It was used as antigen in an ELISA test and compared to that of TSE. Serum samples from 51 symptomatic trichinellosis patients--43 with high antibody levels to TSE, 5 of them with positive biopsy and 8 with low levels of these antibodies--as well as 38 from asymptomatic individuals from the area where the trichinellosis outbreaks had occurred and 43 from apparently healthy individuals from a non-endemic area, 37 from patients with intestinal parasitic infections caused by helminth and protozoan parasites--11 from recurrent and 26 from non-recurrent disease--were analyzed by ELISA using both antigens. The ELISA using p43, 47 Ts L1 detected all trichinellosis patients with high antibody levels as well as 6 out of 8 of those with low antibody levels. All control groups were negative. Therefore, this purified fraction allowed the ELISA to be more specific and sensitive for human trichinellosis diagnosis.
Assuntos
Antígenos de Helmintos/imunologia , Western Blotting , Soros Imunes , Trichinella spiralis/imunologia , Triquinelose/imunologia , Animais , Entamebíase/sangue , Entamebíase/imunologia , Ensaio de Imunoadsorção Enzimática , Helmintíase/sangue , Helmintíase/imunologia , Humanos , Enteropatias Parasitárias/sangue , Enteropatias Parasitárias/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Músculos/parasitologia , Trichinella spiralis/crescimento & desenvolvimento , Trichinella spiralis/isolamento & purificação , Triquinelose/sangue , Triquinelose/parasitologiaRESUMO
Biochemically characterised surface and secreted stage specific antigens were used to analyse the response of humans to infection with Trichinella spiralis. Immuno-coprecipitation and SDS-PAGE of labelled proteins were used to dissect the humoral response of Mexican and European infected sera to each component antigen. The results demonstrate that in man, like laboratory rodents, the response to each protein varies independently. In addition, they permit a more rational approach to the development of improved immuno-diagnostic techniques.