Evaluation of cystatin capture ELISA and sodium metaperiodate antigen treated ELISA in diagnosis of toxoplasmosis

ELISA has become the mainstay for clinical serologic evaluation of toxoplasmosis. One of the major obstacles encountered in the evaluation of ELISA is the false-positive results due to immunological cross-reactions with other parasitic diseases that exhibit some lack of specificity. Evaluation of the diagnostic efficacy of two ELISA techniques: Cystatin capture ELISA and sodium metaperiodate treated antigen-ELISA [SMP-ELISA] compared to conventional ELISA, to detect IgG antibodies for crude T. gondii antigen in sera of toxoplasmosis patients. The study was carried out on 50 individuals categorized into three groups. Toxoplasmosis group included 30 patients confirmed by Sabin-Feldman dye test. Other parasitic diseases group included 10 sera from patients with amoebiasis [2], fascioliasis [2], hydatidosis [3] and schistosomiasis [3]. Control group included 10 healthy individuals. All sera under study were examined for the detection of T gondii IgG by three different ELISA techniques: Cystatin capture ELISA, SMP-ELISA amid conventional ELISA. The diagnostic performance of the three tests was statistically compared. Cystatin capture ELISA gave the best diagnostic results with 96.6% sensitivity, 95% specificity, 96.6% Positive Predictive Value [PPV], 95% Negative Predictive Value [NPV] and 96% diagnostic accuracy. In spite of the lower sensitivity and NPV of SMP-ELISA [86.6% and 82.6%, respectively] than the conventional ELISA, it had higher specificity [95%] and PPV [96.3%]. Cystatin capture ELISA improved the diagnostic performance of conventional ELISA in diagnosis of human toxoplasmosis

Genetic analysis of Blastocystis hominis isolated from symptomatic and asymptomatic human hosts in Egypt

Extensive genomic polymorphism was demonstrated among morphologically identical B. hominis isolates. A genetic diversity would be a powerful tool for identification or classification of B. hominis subtypes. In this study, 14 Egyptian B. hominis isolates were collected, 5 of them were isolated from asymptomatic people whose infections were detected during routine medical check-up and 9 were isolated from patients with gastrointestinal symptoms. Restriction fragment length polymorphism [RFLP] analysis of PCR amplified small-subunit rDNA [SSU rDNA] was used to study genetic diversity of B. hominis isolates by 3 different restriction enzymes [Hin-fl, Rsal and Sau3AI]. Cluster analysis of the riboprint patterns showed 7 distinct genotypes out of 14 B. hominis isolates, 4 were previously reported riboprints and 3 were new ones. The frequency of intestinal symptoms was 64% in Blastocystis cases, Abdominal pain was the most frequent symptom 78% [7/9]. There was no definite correlation between RFLP-banding pattern or genetically distinct genotypes and pathogenecity

Study of N-acetyl-beta-D-glucosaminidase in elderly patients with iron deficiency anaemia

Little is known about renal tubular function in elderly patients with iron deficiency anaemia. To investigate renal tubular function in these patients. To achieve this aim, we compared renal tubular function in 30 asymptomatic non smoker elderly patients [over 65 years] having iron deficiency anaemia [group I] with 15 healthy age and sex matched controls [group II]. Based on their ages, group I patients were subdivided into 2 subgroups; group Ia consisted of 15 iron deficiency anaemia patients >/= 65 - 70 years while group Ib consisted of 15 iron deficiency anaemia patients above the age of 70 years. These patients were diagnosed during routine check up. Patients with known cardiac, hepatic, or renal diseases as well as cancer or other comorbid conditions were excluded from the study. Blood and urine samples were obtained for haematological and biochemical analysis. Urinary N-acetyl-beta-D-glucosaminidase [NAG], as an index of renal tubular function, was estimated by spectrophotometry. Mean urinary NAG levels were significantly higher in iron deficiency anaemia patients of both age groups than in controls [F=85. 77, p<0.001] with significantly higher mean values observed in group Ib than in group Ia patients. No statistically significant difference was observed between mean urinary NAG values in males [19 patients] and females [11 patients] [t= 0.56, p= 0.46]. In elderly iron deficiency anaemia patients of both age groups, urinary NAG values showed statistically significant negative correlation with haemoglobin levels [r=-0.673 and -0.811 at p=0.006 and <0.001 for groups Ia and Ib respectively] and were positively correlated with age [r=0.838 and 0.780 at p<0.001 and <0.001 for groups Ia and Ib respectively]. No correlation was detected between urinary NAG and haemoglobin values in either males or females [r=-0.433, p=0.064 for males and r=-0.077, p=0.821 for females]. There was no correlation between urinary NAG and serum iron, total iron binding capacity [TIBC] or serum ferritin in elderly iron deficiency anaemia patients of both age groups [r=-0.136, 0.328 and -0.070 at p=0.63, 0.232 and 0.805 respectively for group Ia] and [r=-0.229, -0.333 and 0.107 at p=0.414, 0.225 and 0.704 respectively for group Ib]. Elderly patients even with asymptomatic iron deficiency anaemia have impaired renal tubular function. Anaemia may be a contributing factor for renal impairment in such patients that is why it should be early diagnosed and treated even if no clinical disease is immediately apparent

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]

Attempt to improve the diagnostic efficacy of enzyme-linked immunosorbent technique for the serodiagnosis of cystic echinococcosis

ELISA with crude hydatid fluid antigen [CHF] is widely used in the immunodiagnosis of cystic echinococcosis, but cross-reactivity with other helminths, overestimating the true prevalence, represents a great limitation. The present study was carried out to comparatively assess the diagnostic performance of three different ELISA techniques; cystatin capture ELISA, sodium metaperiodate treated antigen ELISA [SMP-ELISA] and conventional ELISA, using CHF antigen to detect lgG antibodies in the serum, aiming to increase the diagnostic efficacy of ELISA technique in detection of hydatid disease. The individuals under study were categorized into 3 groups. Hydatidosis group included 30 hydatidosis patients confirmed by IHAT and abdominal ultrasonograpy, computerized tomography or surgical removal with aspiration and cytological examination. Other parasitic diseases group included 10 sera from patients with; amoebiasis [2], fascioliasis [2], toxoplasmosis [3] and schistosomiasis [3]. Control group included 10 healthy individuals. Sera of all groups under study were examined for the detection of E. granulosus IgG by three different EL1SA techniques; cystatin capture ELISA, SMP-ELISA and conventional ELISA. The diagnostic performance of the three tests was statistically compared. The best diagnostic results were obtained with cystatin capture ELISA. with 96.7% sensitivity. 100% specificity, high positive predictive value [PPV] [100%] and negative predictive value [NPV] [95.2%], 98% diagnostic accuracy [DA], and AUC [0.999] with Cl [95%] [0.99-1.0], followed by SMPELISA, with 90% sensitivity, 95% specificity, PPV [96.4%], NPV [86.4%], DA [92%] and AUC [0.995] with CI [95%] [0.97-0.99], then, the conventional ELISA, with 90% sensitivity, 75% specificity. PPV [84.4%], NPV [83.3%], DA [86%], and AUC [0.962] with Cl [95%] [0.88-0.98]. Both cystatin capture ELISA and SMP-ELISA gave equivalent results for the immunodiagnosis of cystic echinococcosis with AUC1-AUC2 < 0.05. Cystatin capture ELISA and SMP-ELISA are valuable tests for serodiagnosis of human cystic echinococcosis as they improved the diagnostic performance of conventional ELISA

Hyperhomocysteinaemia in renal allograft recipients: could methylenetetrahydrofolate reductase gene polymorphism be a determinant risk factor for chronic allograft dysfunction?

The elevated plasma homocysteine could adversely influence long-term renal graft survival by promoting vascular sclerosis in the kidney allograft. This could be mediated through endothelial dysfunction. A polymorphism C677T in the gene coding for the enzyme methylenetetrahydrofolate reductase [MTHFR] was identified. The aim of the present work was to study the influence of the C677T MTHFR gene polymorphism on total plasma homocysteine and folate levels in renal graft recipients, and its impact on chronic graft dysfunction and the associated endothelial dysfunction. Thirty two stable renal allograft recipients were included in this study [group I] and compared with age and sex matched thirty control subjects [group II]. Plasma homocysteine level, plasma folic acid level, plasma van Willebrand factor [vWF] activity together with endothelial dependent and independent brachial artery vascular responses were done for all subjects. MTHFR genotype was determined by PCR in all renal recipients who were further classified accordingly into 3 subgroups: [group Ia] with homozygous TT type, [group Ib] with heterozygous CT type, and [group Ic] is wild CC type. Renal allograft recipients showed significant higher level of homocysteine as compared to control group [44.42 +/- 32.08 vs 11.62 +/- 2.57 respectively, p<0.001]. There was significant endothelial dysfunction in the transplant group as evidenced by the higher vWF [119.71 +/- 17.71 vs 67.60 +/- 28.65 p<0.001] and poorer endothelial dependent dilatation [EDD] of the brachial artery [7.84 +/- 1.08 vs 12.68 +/- 0.96 p<0.001] as compared to the control group. There was significant negative correlation between plasma homocysteine level and creatinine clearance [r=-0.55, p=0.001], suggesting the deleterious effect of hyperhomocysteinaemia on graft function. The homozygous subgroup [gpIa] showed significant higher level of homocysteine, vWF, lower folic acid, creatinine clearance and EDD as compared to the other two subgroups [gp Ib and Ic]. Our study identified that the presence of hyperhomocysteinemia in combination with unfavorable MTHFR genotypes contributes to an increased risk for development of chronic allograft dysfunction

Impact of experimental duel infections with schistosoma mansoni and echinoccocus granulosus on hepatic histopathology

Experimental duel infection with S.mansoni and E. granulosus was induced in mice to determine their effect on serum nitric oxide [NO] level and accordingly on the sequences of histopathological lesions affecting the liver. The results showed that serum NO level was significantly increased [p

Evaluation of the in vitro effect of Nigella sativa aqueous extract on blastocystis hominis isolates

The effect of Nigella sativa aqueous extract was evaluated against the in vitro growth of 2 different isolates of the intestinal protozoan parasite Blastocystis hominis. Different concentrations [10, 100, 500 microg/ml] of Nigella aqueous extract and metronidazole, an active standard drug for B. hominis, were incubated with B. hominis isolates in culture media at 37°C. Their possible effect on B. hominnis living cell count [LCC] was assessed on Day 1, 3 and 6. The aqueous extract of N. sativa at concentrations of 100 and 500 microg/ml showed a potent lethal effect on both B. hominis isolates, but with different extent. There is no significant difference between the inhibitory effect of N. sativa and metronidazole on the LCC on the 6[th] day. On assessment of living cell rate [LCR] which calculate percentage rate of living cell, N. sativa at 500 microg/ml concentration has a significant inhibitory effect on both isolates. So, it is considered as the most active concentration of Nigella aqueous extract. These results prove that N. sativa aqueous extract could be useful in the treatment of B. hominis

Characterization of specific trichomonas vaginalis target antigens from different isolates by immuno-blotting against hyperimmune rabbit serum

The SDS-PAGE and immunoblot methods were used to identify Trichomonas vaginalis specific target antigen[s]. Ten T. vaginalis isolates, cultured on TYM media, were analyzed by Sodium Dodecyl Sulphate Poly Acrylamide Gel Electrophoresis [SDS-PAGE], revealed 34 distinct bands. Immunoblotting against a hyperimmune rabbit serum, showed 20 reactive bands ranging from 14-205 kDa. There was isolate to isolate variability among 8 isolates, while 2 isolates [6, 7] showed a similar antigenic patterns. Imunoblotting of the isolates showed a total of 20 molecular weight bands, 80% of isolates gave positive immunologic reactions above 100 kDa, while below 100 Kda all the isolates recognized the different molecular weight bands. Out of 20 reacting bands, 5 main bands were detected, 29, 66, 84, 95 and 115 kDa gave positive percent of 60, 60, 60, 90 and 80% among the 10 isolates respectively

Molecular characterization of Egyptian Giardia lamblia isolates

Six Giardia lamblia strains [four from Egypt, one from the USA and one from Sudan] were used to study the phenotypic and genotypic variation in some Egyptian G. Lamblia strains compared with other G. lamblia strains, which may be responsible for the difference in their behavioral characteristics. By using sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] for antigenic study, E1 strain appeared different from the rest of the strains with two bands; one at 121 kDa and the other at 34 kDa, which were not present in other strains, while bands at 130 and 43 kDa were present in all strains, but absent in E1 strain. Another difference between E1 strain and the rest of the strains was obtained by cellulase acetate electrophoresis [CAE], using phosphor-glucomutase [PGM] enzyme, where E1 strain gave a different zymodeme than the other strains. Based on the computerized random amplified polymorphic deoxyribonucleic acid- polymerase chain reaction [RAPD-PCR] analysis, 4 rapdemes were identified; rapdeme 1 contained E1 strain, rapdeme 2 contained E2, E3 and E4 isolates, rapdeme 3 contained Sudan strain and rapdeme 4 contained USA strain. In conclusion, this study revealed a diversity between G. lamblia strains, especially E1 strain, which showed unique characters

preliminary study on the relationship between trichomonas vaginalis and cervical cancer in Egyptian women

The relationship between Trichomonas vaginalis and cancer cervix was investigated by detection of T. vaginalis antibodies in the sera of 48 invasive cervical cancer patients and 100 random age matched female controls using western immunoblot technique. It was found that antibodies to T. vaginalis were detected in sera of 18.75% of cervical cancer patients compared with 5% of the controls. The increase was evident in the age group 40-49 years and of those with squamous cell carcinoma [6/9] and mainly with grades II and III. All the reactive sera of invasive cancer patients reacted strongly with T. Vaginalis surface antigen of about 109.9, 86.1, 56.2, 48.2 and 30 Kda. So, there may be an association between T. vaginalis and the risk of cervical cancer, as there was more than 3-fold increase in the prevalence of T. Vaginalis antibodies in patients with invasive cervical cancer compared with the age matched female controls. This study highlighted the importance of clinically detection of T. vaginalis infection, which is among the factors involved in the genesis and progression of cervical cancer. In addition, its treatment would aid in restricting the rising incidence of this disease

Effect of Schistosoma mansoni egg depostion on multiple low doses streptozotocin induced insulin dependent diabetes

To explore the role of Schistosoma mansoni infection in the prevention of autoimmune mediated insulin-dependent diabetes mellitus, the study examined the effects of multiple low doses of the pancreatic islets beta cell toxin, streptozotocin STZ [40 mg/kg] body weight i.p., given eight weeks post infection period of S. mansoni egg deposition on S. mansoni infected C57BL/6J mice in comparison to non-infected STZ given group. G IV was the control group. Mice treated with STZ [G III] became gradually hyperglycemic reaching the highest level on day 17 post STZ. S. Mansoni infection [G II] significantly reduced the elevation in blood glucose levels from day seven post STZ onwards. Morphologic examination of pancreas on day 21 post STZ revealed that the non-infected STZ [G III] given mice had significantly smaller mean islets area and significantly fewer mean number of beta cells/islets. Pancreatic tissue revealed also focal degeneration in the cells of islets of Langerhans in the non-infected STZ given mice [G III] in comparison to the infected STZ given group [G II], which had much less evident cells degeneration

Evaluation of the effect of a plant alkaloid [berberin derived from berberis aristata] on trichomonas vaginalis in vitro

Berberine is a quaternary alkaloid derived from the plant Berberis aristata having antibanterial, antiamoebic, antifungal, antihelminthic, leishmanicidal and tuberculostatic properties. The effect of berberine sulphate salt on the growth of Trichomonas vaginalis in vitro was compared to the efficacy of metronidazole as a reference drug. Results showed that berberine sulphate was comparable to metronidazole as regards potency with the advantage of being more safe and possible replacement in metronidazole resistant cases

Study of carpal tunnel syndrome [CTS] in uraemic patients on haemodialysis, its relation to B2 microglobulin and arterio-Venous shunt

to assess the prevalence of carpal tunnel syndrome [CTS] in patients with chronic renal failure [CRF] undergoing haemodialysis and to clarify the role of the various factors in its occurrence. Patients and two samples: patients sample: 20 patients with CRF undergoing haemodialysis in the main university hospital in Alexandria divided into two subgroups according to the duration of haemodialysis. subgroup A: less than 5 years and B: more than 5 years. The control sample: ten healthy persons with matched age and sex. All patients were subjected to history taking and neurological examination. Laboratory investigations included blood urea, haemoglobin and serum creatinine, calcium, phosphorus, sodium potassium and beta2-microglobulin concentrations. Electro-physological studies were done using Dantec Cantata machine. Motor and sensory conduction studies, and electromyography were performed to confirm diagnosis of the neuropathy. monoeuropathy in the form of CTS was diagnosed clinically in 55% of cases and electrophysiologicaly in 65% beta2M was significantly increased with prolonged duration of haemodialysis. However, there was no significant effect of age, sex, serum level of beta2M, the duration of haemodialysis or the side of the arterio-venous shunt when studied individually no the occurrence of CTS. However, there was no significant effect of the type of dialysis membranes. the presence of arteriovenous fistula was associated with the occurrence of CTS in a high% age of patients [65%]. Haemodailysis was associated with increased serum level of beta2-M especially in patients with prolonged duration of haemodialysis. No single variable is incriminated in the pathogenesis of CTs and it is more likely that many factors share in its occurrence i.e. it is multi-factorial. Electrophysiological assessment of uraemic patients on haemodialysis revealed subclinical CTS and hence early treatment may result in better prognosis

Hepatitis B vaccination in hemodialysis patients: protective level of HBV antibodies could be achieved by the ordinary schedule?

Hepatits B vaccine is effective in producing protection against hepatitis B virus [HBV] infection in hemodialysis [HD] patients, but the antibody response is variable, so different vaccine regimens were adopted. The aim of the present work is to assess the immune response to HBV vaccine in 67 HD patients receiving 20 micro g of recombinant HBV vaccine / dose for 3 doses, intramuscularly [IM], at 0, 1, and 6- month intervals, compared with 36 medical staff members in the same unit as control receiving the same regimen. The results showed the presence of anti-HBc antibodies in 34 patients and 11 control subjects, those were exculded from the study so as to assess the effect of the vaccine only and not the natural infection. The antibody response is defined as protective when the level of anti-HBs antibodies exceeds or equals to 10 mlU/ml. The response rate was significantly lower in HD patients [51.5%] than in control subjects [96%] [P= 0.0002]. To months after the last dose of the vaccine the difference in seroprotection rate was insigificant being 62.5% in HD patients versus 66.7% in the control [P = 0.89], while after 6 and 12 months, the difference was statistically significant being 56.3%, 33.3% in HD patients versus 100%, and 100% in the control subjects respectively [P = 0.003, 0.006]. On the whole, the levels of anti-HBs, expressed as geometric mean titres and 95% confidence intervals [GMT [95% Cl]], were non-significantly lower in HD patients 125.64 [58.37-269.99] mlU/ml, than controls 239.67 [137.88-416.63] mlU/ml respectively [P = 0.149]. The gender, age, cause of renal failure, HCV co-infection, duration of HD, HD frequency, dialyzer membrane and dialysate buffer did not show a significant influence on the response to HBV vaccine. There were significant positive correlations between the level of anti-HBs antibodies and both serum albumin and predialysis urea [P = 0.00] indicating the improtant role of malnutrition on the poor response to the vaccine in HD patients. From this study it was concluded that the ordinary three-doses regimen is not enough for protection in a good proportion of HD patients, and other regimens should be assessed