Diagnóstico serológico de la esporotricosis mediante el empleo del antígeno de micelio de Sporothrix schenckii sensu stricto / Serological diagnosis of sporotrichosis using an antigen of Sporothrix schenckii sensu stricto mycelium

En este estudio se desarrolló y se evaluó el ensayo por inmunoabsorción ligado a enzimas (ELISA), para la detección de anticuerpos en sueros de pacientes con esporotricosis, para lo cual se empleó un antígeno crudo de Sporothrix schenckii sensu stricto obtenido a partir de la forma micelial. Los sueros positivos para esporotricosis fueron ensayados por otras técnicas serológicas: inmunodifusión doble (IDD) y contrainmunoelectroforesis (CIE). El ensayo fue validado utilizando sueros de otras patologías como histoplasmosis, paracoccidioidomicosis, tuberculosis, leishmaniasis, lupus y sueros de individuos sanos como controles negativos. Se encontró una especificidad de 100 % con las técnicas utilizadas y una sensibilidad del antígeno de S.schenckii sensu stricto, por encima del 98% para IDD, CIE y ELISA. Estos resultados demuestran la alta sensibilidad y especificidad del antígeno de S. schenckii sensu stricto, para el diagnóstico de la esporotricosis, empleando las técnicas de IDD, CIE y ELISA. Los resultados sugieren, que este antígeno podría ser usado en conjunto con otras pruebas convencionales para el diagnóstico diferencial y puede ser útil para monitorizar la evolución de la enfermedad y respuesta al tratamiento.

We developed and analyzed an Enzyme-Linked Immunosorbent Assay (ELISA) in order to detect antibodies in sera from sporotrichosis patients. We used a crude antigen of Sporothrix schenckii sensu stricto, obtained from the mycelial phase of the fungi. Positive sera were analyzed by other serological techniques such as double immunodiffusion (IGG) and counterimmunoelectrophoresis (CIE). The assay was validated by using sera from patients with other pathologies such as: histoplasmosis, paracoccidioidomycosis, tuberculosis, leishmaniasis, lupus and healthy individuals as negative controls. For the Sporothrix schenckii sensu stricto antigen, we found a 100% of specificity by every technique and sensitivity higher than 98% with IDD, CIE and ELISA. Our results show a high sensitivity and specificity for the Sporothrix schenckii sensu stricto antigen, so it can be used for IDD, CIE and ELISA. The results suggest that this antigen could be used in conjunction with other conventional tests for differential diagnosis and may be useful for monitoring the disease progression and response to treatment.

Diagnóstico serológico de tuberculosis pulmonar por la técnica contrainmunoelectroforesis (CIE) / Serodiagnosis of tuberculosis by counterimmunoelectrophoresis (CIE)

Se estudiaron por la técnica Contrainmunoelectroforesis (CIE) 3 grupos de sueros, los cuales incluyeron 56 pacientes con tuberculosis pulmonar, 50 con otras patologías pulmonares y 75 de individuos supuestamente sanos. Se utilizaron en dicha técnica 2 extractos antigénicos celulares crudos de las cepas Mycobacterium tuberculosis H37Ra y Mycobacterium bovis BCG elaborados según Rojas - Espinoza y Quesada - Pascual; se discute la utilidad de la CIE para el diagnóstico serológico de la tuberculosis reportando una sensibilidad de 80,35 por ciento y una especificidad de 93,33 por ciento

Counter immunoelectrophoresis, a modified technique for the identification of blood meals of sandflies collected from Qualyobia governorate, Egypt

Specificity and sensitivity are very important when investigating the blood meals of an insect vector, particularly those having different hosts. Several techniques are used for the identification of the blood meals. Counter immunoelectrophoresis is the technique of choice. The modification was to secure saving of time and least amount of materials. In this paper, a modified technique of the counter immunoelectrophoresis [CIEP] was used for the identification of the blood meal of wild Phlebotomus papatasi collected from different sites in Qalyoubia Governorate. Eight antisera were used. These were human, cat, dog, rat, sheep, goat, bovine, and avian. The results showed that 99.52% of 832 female P. papatasi contained human blood, 0.12% [one female] contained only avian blood, and 0.36% or three females each contained mixed blood of human together with dog blood [one], rat blood [one] or avian blood [one]. P. papatasi is an anthropophilic insect, but in rare cases, it is zoophilic

Diagnosis of active urinary schistosomiasis using homologous adult schistosoma haematobium antigen

The role of antigen prepared from Schistosoma hematobium adult worm [ShAWA, WHO, Geneva] in diagnosis of active urinary schistosomiasis, by counter-immunoelectrophoresis [CIEP], was evaluated by comparison with simpler methods as sedimentation of urinary ova and detection of hematuria by chemical reagent strips. Sixty patients [30 school boys and 30 adults] passing living S. hematobium eggs, as detected, by nucleopore filtration technique [NF], and 30 controls were studied. Results showed statistically significantly higher [P <0.001] eggs counts in school boys than adults. Sensitivities of hematuria detection, ova sedimentation and CIEP were 93.3%, 80%, 50%, 70%, 73.3%, 33.3% in school boys and adults, respectively. It was concluded that direct parasitological examination of urine samples is more superior than CIEP using homologous ShAWA to detect active urinary schistosomiasis both qualitatively and quantitatively. Hematuria detected by chemical reagent strips can identify a high proportion of active urinary schistosomiasis in epidemiological surveys in endemic areas. Nucleopore filtration is more superior than ova sedimentation method for detection of active urinary schistosomiasis

Evaluation of the adult fasciola antigens in immunodiagnosis of human fascioliasis by counter immunoelectrophoresis test

Sera of sixty infected patients with fascioliasis were obtained to evaluate the sensitivity of different antigens from adult Fasciola gigantica [crude, sonicated and purified] by using counter immunoelectrophoresis [CIEP] test for the diagnosis of the infected persons. Sera of twenty normal control persons were collected to evaluate the specificity of different antigens by using C.I.E.P. test. The sensitivity of purified Fasciola antigen was higher [100%] than sonicated [91%] or crude [80%] Fasciola antigen. Fasciola purified antigen gave 100% specificity, while sonicated and crude antigen gave 90% and 75% specificity, respectively. Eosinophilia [11-30%] was recorded in all infected cases with Fasciola. From the results, it may be concluded that purified Fasciola antigen is more sensitive and specific when used in CIEP test. It was recommended to be used where extensive cross reactions are suspected. Also, the eosinophilia may be used as a diagnostic feature of fascioliasis

Antibodies to extractable nuclear antigens in connective tissue disorders in India: prevalence and clinical correlations.

Antibodies to Extractable Nuclear Antigens (ENAs) namely Sm, nRNP, SS-A and SS-B were studied in 397 patients with various connective tissue diseases (CTD), 146 patients with inflammatory polyarthropathies, 16 cases of systemic vasculitides, and 39 normal subjects using counterimmunoelectrophoresis and double immunodiffusion methods. Anti-ENA antibodies were positive in 40.8 percent cases of Systematic lupus erythematosus (SLE) (n = 191), 36.4 percent of overlap CTD (OCTD, n = 44), 27.8 percent of Sjogren's syndrome (n = 18), 10.6 percent of progressive systemic sclerosis (PSS, n = 66) and 2.7 percent of rheumatoid arthritis (n = 111) patients. The correlation of these antibodies with disease features was done. The significant finding was negative association of anti-nRNP antibodies (when present alone) with renal involvement. Anti-Sm antibodies did not correlate with any disease feature. The other associations included correlation of anti-nRNP with pulmonary parenchymal lesions, anti-SS-A with serositis and pulmonary hypertension, and anti-SS-B with myocarditis and recurrent diarrhoea. We conclude that Anti-ENAs may correlate with certain subsets of these diseases but the subject is controversial.

Counterimmunoelectrophoresis test on human Clonorchis sinensis infection.

A counterimmunoelectrophoresis test was used to detect antibodies against the adult worm antigen of Clonorchis sinensis in sera from 70 clonorchiasis patients, 20 uninfected healthy persons and 7 patients infected with other helminths. A constant voltage of 10 V/cm and a running time of 30 minutes was chosen in carrying out detection. Antibody titers of 1, 1:2 and 1:4 were obtained from 35, 21 and 14 clonorchiasis patients, respectively. Significant correlation was observed between worm burden in patients and antibody titer, the higher the antibody titer in patients, the more eggs per gram feces in their stool. Although cross reaction was observed with toxocariasis and angiostrongyliasis in this study, high (100%) sensitivity made it possible to screen the subjects in endemic areas to shorten the survey period.

A comparison of the ELISA with other serological tests in the serodiagnosis of amoebiasis.

The sera of 78 patients with invasive amoebiasis were tested for antiamoebic antibodies by the techniques of enzyme-linked immunosorbent assay (ELISA), indirect haemagglutination (IHA), indirect fluorescent antibody (IFA) and counterimmunoelectrophoresis (CIEP). Results showed that the ELISA compared favourably with IHA and IFA tests in terms of sensitivity and specificity. ELISA, IHA and IFA detected 97.4%, 96.2% and 98.7% of the patients respectively. CIEP was the least sensitive of the 4 serological methods with a sensitivity of 88.5%. The advantages and disadvantages of the 4 serodiagnostic procedures are discussed.