use of ELISA and immunohistochemistry techniques for detection of toxoplasma gondii antigen in tissues of experimentally infected mice

The capability of double antibody sandwich enzyme-linkad immunosorbent assay [ELISA] for detecting antigens of Toxoplasma gondii [T. gondii] in different mice tissue specimens was evaluated in comparison to the immunohistochemistry [IHC] technique. Results proved that tissue antigens were detectable in liver, kidney and mesenteric lymph node [LN] specimens by both methods from the second day of infection, with statistically significant increase in its amount in all organs throughout the period of the study. Using ELISA technique, the highest antigen level was recorded on the second day [0.120 +/- 0.0015] and the fourth day [0.147 +/- 0.0034] of infection in LN specimens, while, the liver showed the highest antigen level at the sixth day post infection [PI][0.165 +/- 0.0066]. On the other hand, using the IHC technique, the highest number of tachyzoites was recorded in LN sections in all studied durations, the second, the fourth and the sixth days PI [1.1 +/- 0.875, 1.6 +/- 1.173 and 3.1 +/- 1.370 respectively]. Thus, sandwich ELISA technique might offer a valuable aid for rapid diagnosis of acute toxoplasmosis in human tissues, and it has proved to be more accurate than IHC technique, since its results was coincided with the pathogenesis of the disease.

rapid diagnostic test for trichomonas vaginalis infection

The applicability of the quantitative buffy coat tube test [QBC] for diagnosis of T. vaginalis was studied. The results of the QBC tube test were compared with that of the wet mount examination in 60 women having genital infection. The cases were selected from Outpatient Clinic of El-Shatby University Hospital. T. vaginalis was identified by QBC tube test in 24 cases out of 60; while with the wet mount smear, it was detected in 21 cases. The sensitivity of the QBC tube test was 100% and the specificity was 92.3%. It was found that the QBC tube test seems to have a role in the routine diagnosis of T. vaginalis

Detection of circulating antigens in blood to evaluate treatment of fascioliasis

This work was designed to assess cure of human fascioliasis after triclabendazole treatment by detection of Fasciola antigen by a modified double antibody sandwich ELISA technique. The results showed that the test detected the antigen in the sera of all studied cases before treatment, while no antigen was detected after treatment. The results suggested that antigen detection provides an accurate tool for diagnosis as well as the assessment of cure

Opportunistic intestinal protozoa in chronic diarrhoeic immunosuppressed patients

The present work aimed to search for the intestinal opportunistic parasites which cause chronic diarrhea. Special stains for each parasite were used to differentiate it easily from the fecal elements obviating the need for diagnostic invasive techniques, especially used in microsporidial infection. The detected parasites were Giardia lamblia [17.7%], as best seen by iron hematoxylin stain and Coccidian oocysts [Cryptosporidia, 13.3% and Isospora belli, 2.2%], as clearly seen by using Ziehl-Neelsen and chromotrope-based stains. The gram positive spores of Enterocytozoon bieneusi were [4.4%] best seen by using chromotrope-based stains; whereas, the diagnosis of Giemsa was failed

Opsonization and intracellular killing of toxoplasma gondii by human mononuclear phagocytes

This study demonstrated that tachyzoites in the presence of antitoxoplasma antibodies with complement were detected in 39 - 58% of monocytes that had phagocytosed them and the mean number of Toxoplasma tachyzoites in this group was significantly low at 1 hour post- infection, while only 10 - 25% of monocytes phagocytosed Toxoplasma tachyzoites in absence of antibodies and complement with significant high number of replicated tachyzoites. This indicated that specific antibodies had a strong opsonizing action. Complement alone was weak in increasing the phagocytic activity

Therapeutic effect of triclabendazole in patients with fascioliasis in Egypt: a preliminary study

10 cases were presented with a variability of clinical manifestations suggesting fascioliasis. The indirect hemagglutination test [IHA] was positive with high eosinophilia. SGOT and SGPT were normal and no signs of heart or kidney diseases were observed. After taking the consent of the patients, triclabendazole, a novel benzimidazole compound was given orally in a single dose of 10 mg/kg body weight. The results revealed that the general condition and all signs of the patients were ameliorated with a drop in the eosinophilic count, which returned to normal after 2-3 months. Being safe, reliable and easy to administer, triclabendazole is ought to be considered for the treatment of human

Elisa Screening for lyme disease in children with chronic arthritis

100 patients aged 6 - 15 years presented to El-Shattby University Children's Hospital with chronic/recurrent arthritis and skin lesions suggestive of erythema chronicum migrans [ECM] were investigated for presence of IgG antibodies against Borrelia burgdorferi flagellum antigen using ELISA test. Four cases yielded positive serum samples which were true positive cases as they showed negative Venereal Disease Research Laboratory [VDRL] test. Ticks collected from domestic animals from the houses of serologically positive patients were found belonging to hard tick, Rhipicephalus sanguineus. Based on clinical ground, only one case was strongly suspected to have Lyme disease due to the presence of the distinctive skin lesion [ECM], while the other 3 cases were not diagnosed till the results of serological test. This study provided serological evidence concerning the presences of Lyme disease in Egypt. Furthermore efforts to identify its prevalence. Various clinical presentations, vectors and animal hosts are warranted

schistosoma mansoni: a comparative study on two cercarial transformation methods

An agar: gelatin plate method, incorporating varying concentrations of linoleate was devised to measure cercarial penetration and transformation in vitro. Schistosoma mansoni cercaria were stimulated over a wide range of linoleate concentrations, while the transformation process occurred over a narrow range. Penetration rates rose gradually until at linoleate concentrations of 0.3 mM and greater, where penetration approached 100%, while transformation just started to occur. The latter then reached maximum [90%] at 3 mM. This optimal concentration of linoleate leading to maximal transformation was then compared with a mechanical transformation method in relation to proteolytic activity and nuclear content. It was found that, proteolytic activity was higher by the chemical than by the mechanical transformation method. The nuclear content decreased by both techniques. This decrease was greater when the mechanical transformation method was applied

Seroprevalence of toxoplasma and cytomegalovirus in complicated pregnancies

Anti-Toxoplasma and anti-cytomegalovirus [CMV] antibodies were sought in 200 pregnant women [100 with complicated pregnancies and 100 matched controls without any hazards] by ELISA test Sixty-five per cent of the women with complicated pregnancies had Toxoplasma antibodies, while 51% had CMV antibodies. Toxoplasma seroprevalence alone was higher in women with intrauterine foetal death [IUFD] [80%]. CMV seroprevalence alone was higher in women with premature labour [44.4%] and women with stillbirth [40%]. Seroprevalence of both Toxoplasma and CMV was clearly higher in women with abortion [55.7%]. Seroprevalence of Toxoplasma and CMV in the control group was 6% and 30% respectively. These data confirm that Toxoplasma and CMV are important causes of complicated pregnancy

Toxoplasmosis: outcome of pregnancy and infant follow-up after prenatal treatment

Twenty-five cases of acquired maternal Toxoplasma infection were the subject of this study. Serological diagnosis was performed using indirect haemagglutination antibody test [IHAT] and enzyme-linked immune sorbent assay [ELISA]. All cases were treated with Spiramycin. After treatment, the level of Toxoplasma antibodies was estimated svery two months using IHAT till antibody level was diminished to be equal or under the diagnostic level. The mean follow-up period was 13.5 months. Out of the 25 cases, 16 cases were continued to term with delivery of 16 live bom babies, 4 cases aborted spontaneously and 5 cases delivered at term but stillbirth. The difference between the IHAT titres before and after treatment was significant in the full-term mothers while there were no statistical significant differences in the aborted and stillbirth cases. Infants' follow-up concerning the level of Toxoplasma antibodies was done every month till antibody level was diminished to be equal or under the diagnostic level. The mean follow-up period was 4.19 months. Serological follow-up of infants after birtti showed a statistical significant difference between the titres at different months and the drop of the titre started at the second month after birth

Intestinal parasitic infection among sewage workers in Alexandria, Egypt

This research was conducted to study the risk of intestinal parasitic infections among sewage workers. A hundred sewage workers in Alexandria and another hundred as control group not exposed to sewage were selected for this research. The results indicated that sewage workers were at a higher risk of developing intestinal parasitic infection, especially Ascariasis and Amoebiasis. Nail examination revealed that sewage workers were at a higher risk of infection by Entrobius vermicularis

Study of antibody titres in human toxoplasmosis and its relation to specific treatment

The percentage distribution of toxoplasmosis among 502 clinically suspected cases was found to be 23.9%. The highest percentage was that of the occular cases, followed by obstetric positive cases, patients with cervical lymphadenopathy, then cerebral and fever groups. Among the obstetric positive cases with toxoplasmosis, highest positivity rate was found in cases with history of stillbirths, then those, with premature births, followed by patients with abortions; sporadic more than habitual abortion. After treatment with pyrimethamine-sulfadiazine, negative results started to appear in the seventh month using IHAT and ELISA, and in the ninth month using IFAT. After treatment with spiramycin, negative results were detected in the ninth month by IHAT and ELISA, and in the eleventh month onwards using IFAT. Comparisons of the sensitivity and specificity of IHAT IFAT and ELISA in diagnosing toxoplasmosis of the examined cases revealed that IFAT can be considered the most sensitive and specific

attempt to induce immunity against toxocara canis infection in albino mice parasltological studies

This study aimed at preparation and fractionation of the crude second stage Toxocara canis larval antigens and the determination of the optimal dose of the crude and fraction that gave the highest percentage reduction in the larval counts. The crude antigen was prepared from the second stage larvae after hatching of the cultivated larvated eggs obtained from the uteri of adult worms collected from 250 stray puppies. The antigen doses given individually in each group with a total dose of 12.5, 25, 50 and 100 mg protein, respectively. The crude antigen was further fractionated by column chromatography using Sephadex G-200. The dose of each fraction was calculated from the crude optimal dose according to their proportion in the elution volume. All immunized mice were challenged two weeks after the last imunizing dose by 800 larvated eggs. Brain larval count was done 30 days post-challenge to determine the optimal dose giveng the highest percentage reduction. It was found that a total dose of 12.5 mg protein of the crude antigen and the fourth fraction gave the highest percentage reduction in the larval count. These two antigens were further used to study the effect of immunization on humoral antibody responses. This was done by determination of the serum antibody level by the indirect fluorescent antibody test 3, 7, 15, 30 and 60 days post-infection. The antibody responses early and higher in the immunized groups than those of the controls. The maximum antibody level was reached at 15 days post-infection in all groups

Interleukin-2 level in mice infected with toxoplasma gondii

This work described the altered Interleukin-2 levels [IL-2] in mice infected with a virulent strain of Toxoplasma gondii and some possible mechanisms responsible for the alteration. To investigate this point, the effect of Toxoplasma infection was assessed shortly one week and then one month after invasion. Interleukin-2 production by stimulated Con A was reduced one week after infection and markedly depressed one month after infection with respect to normal controls. Unstimulated spleen cells from infected mice produced more IL-2 in vitro than stimulated cells. So, it can be concluded that IL-2 is one of the causes of immune suppression of Toxoplasmosis

pathogenesis of giardia lamblia among Saudi children

The pathogenicity of giardiasis was studied among children of rural areas in the surrounding of Jeddah. Stools showing Giardia lamblia were examined macroscopically and microscopically to detect evidences of functional intestinal disturbances. The load of Giardia lamblia infection was assessed among them. The results showed that in this group of children giardiasis was not commonly associated with evident functional intestinal disturbances

Prevalence of parasitic infections in a rural community in Saudi Arabia

The prevalence of parasitic infections among 680 subjects living in a rural and suburban community near Makkah was studied [About half the population in such communities harbour one or more parasitic infections]. Among the most important environmental factors shown to affect the prevalence of intestinal parasites in this study are the level of environmental sanitation, the area of residence [whether urban or rural], family size, and sex. On the other hand, age appeared to play a minor or no role

Effects of nematode larvae [trichinella spiralis and toxocara canis] on B-glucuronidase enzyme in experimentally infected mice

Experimental infection of mice with Trichinella spiralis and Toxocara canis larvae increased the activity of B-glucuronidase enzyme in the liver and splenic homogenates in both infections and in the kidney homogenates in the former infection. This rise in B-glucuronidase indicates tissue damage which was attributed mostly to immunological nature