Identification and evaluation of antigenic epitopes of total soluble extract and adult worm vomit for immunodiagnosis of human fascioliasis

Fascioliasis is an important worldwide herbivores infection with increasing evidence of prevalence as a human disease. There is an important necessity to develop sensitive and specific diagnostic tools for early treatment and to avoid complications. This study is directed to identify, characterize and evaluate the immunodiagnostic potential of the total soluble extract [TSE] and the adult worm vomit [AWV] antigens of worm using immunoblotting of total IgG and IgG4. Sixty individuals, 29 positive for fascioliasis by IHAT [Group I], 15 positive for other parasitic infections [Group II] and 16 normal controls [Group III], were included. Both TSE and AWV antigens were analyzed by SDS-PAGE. All sera were immunoblotted against the two antigens using total IgG and IgG4. Total IgG immunoblotting using TSE antigen at 62 kDa gave sensitivity, specificity, positive value predictive [PVP] and accuracy of 72.5%, 100%, 100% and 82.2%, respectively. These values were 62.5%, 100%, 100% and 81.2%, respectively with AWV antigen at 56 kDa. IgG4 immunoblotting using TSE antigen gave sensitivity, specificity, PVP and accuracy of 100%, 93.3%, 96.6% and 97.8%, respectively at 60 kDa, while AWV antigen gave no reaction with IgG4 antibody. Total IgG immunoblotting using TSE antigen at 62 kDa and AWV antigen at 56 kDa proved to be valuable as diagnostic tests for human fascioliasis

single-step immunochromatographic lateral-flow assay for detection of giardia lamblia and cryptosporidium parvum antigens in human fecal samples

REDA Quick immunochromatographic lateral-flow assay was evaluated for diagnosis of giardiasis and cryptosporidiosis as compared to the [gold standard] stool examination. Of the 300 specimens were examined by microscopy of direct wet films, concentrated sediments, modified trichrome and modified Ziehl-Neelsen stained slides, 35 samples of Giardia, ten Cryptosporidium, 35 of other parasites, and 20 negative controls were selected for RIDA Quick test examination. All the samples that gave discrepancy results were retested by the centrifugation prior to preparation for the permanent stained smear. The sensitivity, specificity, positive predictive value [PPV] and negative predictive value [NPV] of RIDA quick test for Giardia were 91.6%, 98.4%, 97% and 95.4% respectively, while that of the microscopic stool examination were 94.5%, 100%, 100% and 96.9%: The sensitivity, specificity, PPV and NPV of RIDA quick test for Cryptosporidium was 91.6%, 100%, 100% and 98.8% respectively, while that of the microscopic stool examination were 83.3%, 100%, 100% and 97.7%

Detection of molecular markers of toxoplasmosis among Egyptian patients with miscarriage using avidity IGG-ELISA and western blotting

A total of 54 miscarriage patients were divided into 3 groups. GI: 10 toxoplasmosis patients with +ve IgM-ELISA; GII 24 toxoplasmosis patients with +ve IgG-ELISA, and G III: 20 non-toxoplasmosis cross-matched females as a control. All groups were subjected to IgG-avidity ELISA and IgG-avidity immunoblotting. Avidity Indices [AI] by ELISA ranged from 22.6% to 73.3% in GI and from 9.6%-75.6% in GII AI were high [>40%] in 3 [30%] patients in G I and in 8 [33.3%] patients in G II. Sera of GI recognized the 20, 28,32,60,93 and l00 Kda bands with 55% reduction in the 38 and 60 Kda bands after treatment with 6 M urea solutions. Sera of G II recognized the 20, 28, 32, 38, 45, 95-97 and 106 Kda bands. There was 12.5%, 16.6% and 16.7% reduction in the 20, 32, and 106 Kda bands, respectively, after urea. The 38 and 60 Kda bands were identified as good diagnostic markers for the recent toxoplasmosis infection [GI]. The 20, 32 and 106 Kda bands were good markers of high avidity antibodies during the chronic toxoplasmosis [GIl]

Immunopathogenic role of IgG antibody and rantes in house dust mite-induced chronic bronchitis

Based on immunological and clinical examinations, 21 patients were diagnosed as having house dust mite [HDM]-induced chronic bronchitis and classified into three groups according to the clinical presentation of the disease; stable bronchitis, exacerbated bronchitis and asthma on top of bronchitis. Using ELISA, the levels of serum anti-Dermatophagoides farinae and anti-D. pteronyssinus IgG antibodies and plasma RANTES [regulated upon activation, normal T-cell-expressed and secreted; a chemokine with attractive and activator role for eosinophils] were measured in correlation to serum eosinophil cationic protein [ECP, a marker of eosinophil activation and degranulation measured by chemiluminescent immunometric technique]. Using immunoblotting, IgG binding components of D. farinae and D. pteronyssinus were determined providing a clue for diagnosis of HDM-induced chronic bronchitis. Significant higher levels of anti-D. farinae and anti-D. pteronyssinus IgG antibodies and RANTES were found in asthmatic group, followed by exacerbated chronic bronchitis in comparison to stable bronchitis and control groups. ECP level correlated significantly with IgG and RANTES levels in exacerbated bronchitis and asthmatic groups. The results provided evidence that over expression of IgG and RANTES plays a crucial role as mediator in immunopathogenesis of HDM-induced chronic bronchitis and as marker of the immunological changes likely responsible for progression of bronchitis to asthma in HDM-sensitive patients, yet RANTES seemed to be an early indicator. Definition of the immunopathogenic role of IgG and RANTES in HDM-induced bronchitis should enable the manipulation of the critical immune response in the hope of establishing new therapies. D. farinae and D. pteronyssinus antigenic bands at >205 and 205 KDa, respectively, considered together showed 71.4% sensitivity in diagnosis of HDM induced chronic bronchitis and 100% specificity by immunoblotting

Avidity of immunoglobuling antibody response to the different antigenic fractions of soluble Schistosoma mansoni adult worm antigen preparation [SWAP] using Avidity Immmunoblotting assay

Human serum samples from 25 acute schistosomiasis patients, 20 chronic cases and 15 normal healthy controls were analyzed by IgG avidity enzyme linked immunosorbent assay [ELISA] and IgG avidity immunoblotting assay. Using avidity ELISA, a pronounced overlap of avidity index values was observed between acute and chronic infections with a range of uncertainty [0.86-1], which was encountered in both groups. Using avidity immunoblotting assay, antigenic bands at >116, 84, 48, 40 and 34 kDa were exclusive for the acute phase. From these bands, 34 kDa was recognized mostly by low-avidity antibodies and showed a high sensitivity [96%] and specificity [100%], making it an optimal marker for the acute phase. 40 kDa band was recognized mostly by high-avidity antibodies, even during acute infection. Bands of 80, 70, 42, 36, 30 and 26 kDa were exclusive for the chronic phase. Only 70 kDa band was recognized by high-avidity antibodies, yet with low sensitivity [35%] that limits its use as an optimal marker for the chronic infection. Meanwhile, 70 and 40 kDa bands, recognized by high-avidity antibodies, are considered as potential vaccine antigen candidates

Evaluation of semi - quantitative PCR and IgG and IgM Elisa in Diagnosis of toxoplasmosis in females with miscarriage

This study was performed on 100 female [age 20-42 years] patients classified into group I, 40 patients presented with abortion in the first trimester; group II, 33 patients with abortion in the second trimester and group III, 27 patients with intrauterine fetal death [IUFD]. The positive percentages of semi-quantitative PCR and both IgG and IgM ELISA were 38% and 35%, respectively. Ten cases out of 38 were positive for toxoplasmosis by both PCR and ELISA-IgG, while 5 cases were positive by both PCR and ELISA-IgM. Sixteen cases out of 38 PCR positive cases were positive by both ELISA IgG and IgM. Sensitivity and specificity of both ELISA IgG and IgM were 81.57% and 93.54%, respectively. False negative results by ELISA were found in 7 out of 38 positive toxoplasmosis cases detected by semi-quantitative PCR. Three out of those seven cases with false negative by ELISA were detected with a trophozoite copy load of 101 trophozoite/mL in the blood sample by semi-quantitative PCR

Avidity IgG: Diagnosis of primary toxoplasma gondii infection by indirect immunofluorescent test

In the present study, 18 serum samples from toxoplasmosis patients with different clinical manifestations were classified into GI, nine patients suspected of having recent T. gondii infection and G2, nine patients suspected of having distant latent infection. 80% of patients with low IgG avidity antibody were positive by IgG avidity- indirect immunofluorescent antibody test [IFAT] and by IgM-ELISA for toxoplasmosis. However, 20% positive by low IgG avidity-IIFAT, were IgG-ELISA positive for toxoplasmosis. 15.4% of patients with high IgG avidity antibody were positive by high IgG avidity-IIFAT and by IgM- ELISA; but 61.5% positive by high IgG avidity-IIFAT were positive by IgG-ELISA and 23.1% positive by high IgG avidity IFAT were positive with both IgM and IgG ELISA

Nosocomial sources of cryptosporidial infection in newly admitted patients in Ain Shams University Pediatric Hospital

The present study included 104 patients and 27 contacts. Both were subjected to stool examination by modified Ziehl-Neelsen [MZN] stain. ELISA for the detection of cryptosporidium Ag in stool was further carried out for patients developed diarrhea after admission [12], patients admitted with diarrhea [17] and for contacts. Twenty-four water samples were collected from Pediatric Hospital and examined by MZN and ELISA. The results showed that 11.5% of the examined cases developed diarrhea after admission, 8% of them were positive for cryptosporidiosis by both MZN and ELISA. 17% of the contacts were negative by MZN stain. ELISA was found to be 100% sensitive and 94.1% specific. Water samples were found to be negative for C. parvum