RESUMO
Examination of anticardiolipin antibodies (ACAs) in the sera of patients clinically diagnosed with chronic fatigue syndrome (CFS) using an enzyme-linked immunoassay procedure demonstrated the presence of immunoglobulin M isotypes in 95% of CFS serum samples tested. The presence of immunoglobulin G and immunoglobulin A isotypes were also detected in a subset of the samples. Future studies will focus on elucidating whether alterations to mitochondrial inner membranes and/or metabolic functions play a possible role in the expression of ACAs.
Assuntos
Anticorpos Anticardiolipina/sangue , Síndrome de Fadiga Crônica/imunologia , Isotipos de Imunoglobulinas/imunologia , Adulto , Idoso , Cardiolipinas/imunologia , Ensaio de Imunoadsorção Enzimática , Síndrome de Fadiga Crônica/patologia , Feminino , Humanos , Imunoglobulina M/imunologia , Masculino , Pessoa de Meia-Idade , Mitocôndrias Hepáticas/imunologiaRESUMO
A sandwich enzyme-linked immunosorbent assay was developed to detect ciguatoxin (CTX) in fish tissue. The assay utilizes two antibodies, chicken immunoglobulin Y specific to the ABCD domain of CTX and a mouse monoclonal immunoglobulin G-horseradish peroxidase conjugate specific to the JKLM domain of CTX. The sensitivity, working range, cross reactivity, accuracy, precision, and reproducibility were examined.
Assuntos
Galinhas/imunologia , Ciguatera/diagnóstico , Ciguatoxinas/análise , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulinas/imunologia , Venenos/análise , Alimentos Marinhos/análise , Animais , Anticorpos Monoclonais/análise , Bioensaio/métodos , Ciguatoxinas/imunologia , Reações Cruzadas/imunologia , Camundongos , Neuroblastoma , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Células Tumorais CultivadasRESUMO
The applicability of a new enzyme-linked immunoassay (ELISA) for detecting ciguatoxin (CTX) in fish tissue was evaluated by testing three fish species commonly implicated in ciguatera fish poisoning in Hawaii. A total of 164 individual almaco jack (Seriola rivoliana) and greater amberjack (S. dumerili) and a total of 175 individuals of the blue-spotted grouper (Cephalopholis argus) were caught at various locations in the Hawaiian Islands. Muscle tissue from each individual was assessed for the presence of CTX using two methods: a semi-quantitative ELISA that was recently developed for detecting picogram levels of CTX in fish extract and a neuroblastoma (NB) cell assay commonly used to screen for marine toxins in fish. Results of the tests were highly correlated, with the ELISA indicating the presence of CTX in 9.4% of all fish samples, and the NB assay indicating toxicity in 6.8% of the fish samples. We conclude that the ELISA produces reliable and accurate results that are consistent with those provided by the accepted NB assay and that the ELISA has potential for future applications in screening fish populations for CTX.
Assuntos
Ciguatera/diagnóstico , Ciguatoxinas/farmacologia , Ensaio de Imunoadsorção Enzimática/métodos , Contaminação de Alimentos/análise , Alimentos Marinhos/análise , Animais , Bioensaio , Sobrevivência Celular/efeitos dos fármacos , Ciguatoxinas/análise , Monitoramento Ambiental/métodos , Camundongos , Músculo Esquelético/química , Neuroblastoma/tratamento farmacológico , Neuroblastoma/patologia , Perciformes , Reprodutibilidade dos Testes , Células Tumorais CultivadasRESUMO
This study examined 328 CFS sera in a study with 17 CCFP, 8 Gulf War Veterans (GWV), 24 Prostate Cancer (PC), and 52 normal sera in the modified Membrane Immunobead Assay (MIA) procedure for CTX. Three hundred and twenty-eight CFS patients' sera were examined by the modified MIA with purified MAb-CTX and 91.2% gave a titre > or =1:40. 76% of the 17 CCFP sera samples and 100% of the 8 GWV sera samples also had a titre > or =1:40. 92.3% of 52 normal sera showed titres of 1:20 or less, while 4 gave titres of > or =1:40. In addition, 41 sera were examined for Anti-Cardiolipin (aCL) by a commercial ELISA procedure with 87.8% demonstrating IgM, IgM+IgA, or IgM+IgG aCL antibodies. These results showed mostly the IgM aCL antibody alone in the sera samples. In addition, 41 serum samples were examined for aCL, with 37 showing positive for aCL, representing 90.2% positive for the three disease categories examined: CFS, CCFP and GWV. Examination for antiMitochondrial-M2 autoantibody (aM-M2) in 28 patients (CFS (18), CCFP (5), and GWV (5)) was negative for aM-M2. Inhibition analysis with antigens, CTX, CFS "Acute Phase Lipids", commercial Cardiolipin (CL) and 1,2-Dipalmitoyl-sn-Glycero-3-[Phospho-L-Serine] (PS) and antibodies, MAb-CTX and aCL from patients' serum show that the phospholipids in CL and CTX are antigenically indistinguishable with antibodies MAb-CTX and CFS-aCL. Preliminary chemical analyses have shown the lipids to be phospholipids associated with CL of the mitochondria. We designate this "Acute Phase Lipid" comparable to "Acute Phase Proteins" (C-reactive protein (CRP) and Serum Amyloid A (SAA)) in inflammatory conditions.