RESUMO
A total of 70 serum samples from heparin-induced thrombocytopenia (HIT II) patients, non-HIT patients and healthy subjects, respectively, were studied for the presence of antiheparin/PF4 antibodies. Two enzyme-linked immunosorbent assay (ELISA) assays were compared with the particle gel immunoassay (PaGIA). Beads of the PaGIA kit were also used to evaluate the feasibility of flow cytometric detection of antiheparin/PF4 antibodies in patient samples. Experiments have shown that all samples found positive by ELISA and PaGIA, were also positive when analysed by flow cytometry by an indirect test using the high-density particles coated with heparin/PF4 complexes and a second step fluorescein isothiocyanate (FITC) antihuman immunoglobulin (Ig)G reagent. The procedure was easy to perform, repetitive and beads were promptly visualized by physical parameters, with a very low background. In conclusion, the results of this study suggest that flow cytometry is a reliable method for the detection of antiheparin/PF4 antibodies.
Assuntos
Autoanticorpos/sangue , Heparina/imunologia , Fator Plaquetário 4/imunologia , Trombocitopenia/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática/normas , Feminino , Citometria de Fluxo/normas , Heparina/efeitos adversos , Humanos , Isotipos de Imunoglobulinas/sangue , Testes de Fixação do Látex/normas , Masculino , Microesferas , Pessoa de Meia-Idade , Trombocitopenia/induzido quimicamente , Trombocitopenia/imunologiaAssuntos
Membrana Eritrocítica/efeitos dos fármacos , Isoanticorpos/sangue , Papaína/farmacologia , Gravidez/imunologia , Sistema do Grupo Sanguíneo Rh-Hr/imunologia , Citotoxicidade Celular Dependente de Anticorpos , Teste de Coombs , Eritroblastose Fetal/epidemiologia , Eritroblastose Fetal/etiologia , Membrana Eritrocítica/imunologia , Feminino , Humanos , Incidência , Recém-Nascido , Gravidez/sangue , Estudos Retrospectivos , Imunoglobulina rho(D)RESUMO
We performed a flow-cytometric analysis of the expression of D antigen in D+W samples (previously termed Du). We also analysed a series of D-positive and D-negative (cde phenotype) samples to obtain positive and negative controls, respectively. The evaluation was carried out by immunofluorescence and the intensity of positivity was expressed as mean channel value (MCV) of fluorescence. Results demonstrated that D+W samples have lower expression (less than 1 log) than D-positive cases (p > 0.001, Student t test), while cde samples show the same MCVs as negative controls. Moreover, it was also possible to set a grading of D antigen expression and to analyze cases difficult to assess by agglutination only.
Assuntos
Tipagem e Reações Cruzadas Sanguíneas/métodos , Citometria de Fluxo , Sistema do Grupo Sanguíneo Rh-Hr/análise , Tipagem e Reações Cruzadas Sanguíneas/instrumentação , Testes de Hemaglutinação , Humanos , Fenótipo , Sensibilidade e EspecificidadeRESUMO
The persistence of anti-HIV-1 antibodies in bloodstains has been studied by ELISA and Western Blot (WB) analysis. The immunoblot technique was found to be specific and more sensitive and the antibodies could be detected in 0.7 mg of bloodstains for up to 6 months. The authors emphasize the importance of such an investigation on non-genetic markers in individual diagnosis for forensic purposes.
