Comparison of the multi-epitope recombinant antigen DIPOL and hydatid fluid for the diagnosis of patients with cystic echinococcosis.

The use of serological tests containing multiple immunodominant antigens rather than single antigens have the potential to improve the diagnostic performance in Cystic Echinococcoses (CE) as a complement tool to clear the inconclusive imaging data. Here, we comparatively evaluated the diagnostic value of Hydatid Fluid (HF) and the recently described recombinant multi-epitope antigen DIPOL in IgG-ELISA in a clinically defined cohort of CE patients. The serum samples from 149 CE patients were collected just before surgical or Percutaneous- Aspiration- Injection- Reaspiration (PAIR) procedures. Additionally, serum samples of patients with other parasitic infections (n=49) and healthy individuals (n=21) were also included in the study as controls. To investigate the association between the genotype of the parasite and DIPOL, cyst materials from 20 CE patients were sequenced. In terms of overall sensitivity, HF was higher than DIPOL (82.55%,78.52%, respectively). However, while the sensitivity of HF was higher than DIPOL in patients with active and transitional cysts (83.3%, 75.4%, respectively), sensitivity of DIPOL in inactive cysts was higher compared to HF (95.6%, 78.3%, respectively). The sensitivity of DIPOL depending on cyst stage was statistically significant (P= 0.041). In terms of specificity, DIPOL was found to be better than HF (97.71%, 91.43%, respectively). By genotyping, the majority of 20 patients showed G1 genotype (80%). All patients harboring G3 and G1/G3 cyst genotypes were positive with both antigens, while 87.5% of patients with G1 genotype were seropositive with HF and 75% with DIPOL. The overall sensitivity and high specificity of DIPOL suggest that this recombinant protein containing immunodominant epitopes is a potential substitute for the HF by serological tests for the diagnosis of CE.

Comparison of the New Multiepitope Recombinant Peptide Antigen with Cyst Fluid Antigen in the Follow-up of Patients with Cystic Echinococcosis

Objective: The follow-up of patients with cystic echinococcosis (CE) offers the opportunity of evaluating the prognosis of the infection as well as detecting relapse. This study aimed to evaluate the performance of the new multiepitope recombinant peptide (recDipol) antigen in the follow-up of CE patients treated by surgery or percutaneous aspiration injection respiration. Methods: A total of 137 blood samples from 28 patients were evaluated by IgG-ELISA method using recDipol and hydatid fluid (HF) antigens. The patients were simultaneously checked for recurrence by ultrasonography. Results: The seropositivity rate of the 28 patients varied considerably during the follow-up. When the first blood of the patients was evaluated, 4 (14.28%) were seronegative by HF-ELISA and 9 (32.14%) were recDipol-ELISA. During the entire follow-up, only 1 (3.5%) and 6 (21.4%) patients were seronegative by HF-ELISA and recDipol ELISA, respectively. Conclusion: We found that recDipol did not perform as expected in the follow-up due to the higher number of seronegative patients compared to HF-ELISA in the first blood and during the entire follow-up. Our results suggest that imaging methods are gold standards in the diagnosis and that, in parallel, longer-term patient follow-up is required with recombinant antigens that have an improved diagnostic performance.

[Evaluation of the diagnostic value of the ELISA tests developed by using EgHF, Em2 and EmII/3-10 antigens in the serological diagnosis of alveolar echinococcosis]. / Alveoler ekinokokkozun serolojik tanisinda EgHF, Em2 ve EmII/3-10 antijenleri ile hazirlanan ELISA yöntemlerinin tanisal etkinliginin degerlendirilmesi.

Alveolar echinococcosis (AE), caused by larva stage of Echinococcus multilocularis, is one of the lethal parasitic diseases of man and a major public health problem in many countries in the northern hemisphere. When the living conditions and habits in Turkey were considered in terms of relation with the life cycle of the parasite, it was suggested that AE has been much more common than reported mainly from the Eastern Anatolia region of Turkey. Since in vitro serologic diagnosis tests with high specificity for AE have not been used in our country, most of the cases with liver lesions were misdiagnosed by radiological investigations as malignancies. The aim of this study was to evaluate the diagnostic value of the in-house ELISA methods developed by using three different antigens (EgHF, Em2, EmII/3-10) in the serological diagnosis of AE. The study samples included a total of 100 sera provided by Bern University Parasitology Institute where samples were obtained from patients with helminthiasis and all were confirmed by clinical, parasitological and/or histopathological means. Ten samples from each of the cases infected by E.multilocularis, E.granulosus, Taenia solium, Wuchereria bancrofti, Strongyloides stercolaris, Ascaris lumbricoides, Toxocara canis, Trichinella spiralis, Fasciola hepatica and Schistosoma haematobium were studied. In the study, EgHF (E.granulosus hydatid fluid) antigens were prepared in our laboratory from the liver cyst fluids of sheeps with cystic echinococcosis, however Em2 (E.multilocularis metacestode-purified laminated layer) and EmII/3-10 (E.multilocularis recombinant protoscolex tegument) antigens were provided by Bern University Parasitology Institute. Flat bottom ELISA plates were covered with EgHF, Em2 and EmII/3-10 antigens in the concentrations of 2.5 µg, 1 µg and 0.18 µg per well, respectively, and all sera were tested by EgHF-ELISA, Em2-ELISA and EmII/3-10-ELISA methods. For each tests, the samples which were reactive above the cut-off value (mean OD of negative controls+2 SD) were accepted as positive. The sensitivity of the ELISA tests performed with EgHF, Em2 and Em2II/3-10 antigens were estimated as 100%, 90% and 90%, respectively, whereas the specificity were 63%, 91% and 91%, respectively. When Em2-ELISA and EmII/3-10-ELISA tests were evaluated together, the specificity increased to 96%. Our data indicated that the highest sensitivity (100% with EgHF-ELISA) and specificity (96% with Em2-ELISA + EmII/3-10-ELISA) for the serodiagnosis of AE can be achieved by the combined use of the ELISA tests with three different antigens. It was concluded that the early and accurate diagnosis of AE in our country which is endemic for that disease, could be supported by the use of highly specific serological tests such as Em2-ELISA ve EmII/3-10-ELISA contributing radiological data.

Evaluation of purified antigen fraction in the immunodiagnosis of cystic echinococcosis.

Echinococcus granulosus antigen B fraction (AgB) was evaluated for its prognostic value in the serological follow-up of cystic echinococcosis (CE), compared to crude hydatid fluid (HF) as well as soluble somatic Echinococcus multilocularis antigen (Em). The sensitivity and specificity of enzyme-linked immunosorbent assays (ELISA) were compared examining 177 sera from patients with different clinical courses and outcome of CE and with other parasitic infections. AgB-ELISA in comparison to confirmed cystic echinococcosis has 96.4% sensitivity and 97.2% specificity, with 93.1% positive predictive value and 98.6% negative predictive value. The HF-ELISA was more sensitive than the AgB-ELISA, but its specificity was lower. Our results indicate that AgB-ELISA was more satisfactory for seroconfirmation of acute echinococcosis than HF-ELISA. Moreover, the AgB-ELISA has a potential key role in control measures implemented in patients undergoing surgery. In sera of patients more than 3 months after the treatment, no antibody response to antigen B was detected; however, with conventionally used HF-ELISA, they were still positive. The AgB is recommended for the confirmatory diagnosis. AgB-WB allocated low background with typical "triplet" bands at 8-12-16 kDa.

The investigation of congenital toxoplasmosis in a tertiary care hospital in Turkey.

OBJECTIVE: To elucidate the prevalence of congenital toxoplasmosis in Turkey. METHODS: This study was conducted in the Department of Parasitology, Faculty of Medicine, Ege University, Izmir, Turkey during the period of 2006 and 2007. Venous blood before delivery and cord blood during delivery were collected from 138 women, and we observed the presence of immunoglobulin (Ig)G and IgM antibodies by Enzyme Linked Immunosorbant Assay (ELISA), western blotting, and other serologic tests. RESULTS: The combination of ELISA and western blotting have the greatest sensitivity among the serological techniques used. The results of the cord and venous sera were comparable with no significant difference, except for one sample. CONCLUSION: Data obtained showed that early characterization of IgG antibodies synthesized by congenitally infected newborn is important. This preliminary study sets an example to the studies that would help both in determining the incidence of congenital toxoplasmosis by screening larger populations and in preventing sequels by early diagnosis.

Parasitic zoonotic diseases in Turkey.

Zoonoses and zoonotic diseases are becoming more common and they are now receiving increased attention across the world. Zoonotic parasites are found in a wide variety of protozoa, cestodes, nematodes, trematodes and arthropods worldwide and many zoonotic parasites have assumed an important role. The importance of some parasitic zoonoses has increased in recent years due to the fact that they can be agents of opportunistic infections. Although a number of zoonotic parasites are often found and do cause serious illnesses in Turkey, some are more common and these diseases are more important as they cause serious public health problems, such as leishmaniasis, toxoplasmosis, cryptosporidiosis, echinococcosis, trichinellosis and toxocariasis. Information on these zoonotic diseases is provided here as these are the most important zoonotic parasitic diseases in Turkey.

[A study of sister chromatid exchange (SCE) in patients with liver cystic hydatid disease who have been treated with albendazole.]. / Karaciger kist hidatigi tedavisinde albendazol kullanan hastalarda kardes kromatid degisimi (SCE) çalismasi

A total of 21 patients including 13 females and 8 males who presented at the Microbiology and General Surgery Departments of the Atatürk Research and Training Hospital with complaints of liver cystic hydatid disease have been included in this study. Albendazole was administered to these cystic hydatid patients before and after surgery. The aim of this study was to investigate the probable genotoxic effects of albendazole. SCE testing was applied to the blood samples taken from patients after the treatment and the mutagenic effect of albendazole was investigated by comparison with the control group. Student t- test was used for the statistical analysis of the results.

Past to present: echinococcosis in Turkey.

Echinococcosis is a zoonotic infection caused by Echinococcus spp. and is one of the most important helminthic diseases worldwide. Two forms of echinococcosis occur in Turkey, i.e. Echinococcus granulosus and E. multilocularis. The life cycle of E. granulosus is predominantly in dogs and sheep, and most sheep farmers in Turkey keep a dog or two. Stray dogs are numerous and prevention or treatment of infection in these dogs is very difficult. Cystic echinococcosis (CE) occurs throughout Turkey whereas alveolar echinococcosis (AE) predominantly occurs in the eastern Anatolian region of the country. Both CE and AE are known to be endemic but few surveys have been performed. Most data on human CE and AE have been collected from hospital records. The first reference of echinococcosis, "Kyste hydatique multiloculaire", in Turkey dates as far back to 1872 by an Ottoman Physician, C.R. Katibian. The results of the first Turkish study on E. granulosus were published in 1928. According to Ministry of Health records, 21303 patients had operations to treat or confirm CE in the period 1987-1994 which corresponds to approximately 2663 patients per year. The estimated surgical case rate of CE is 0.87-6.6 per 100000 in Turkey. The prevalence of E. granulosus infection in dogs in Turkey is between 0.32 and 40% and varies widely with geographical location. The reported prevalence of CE in domestic animals in Turkey has ranged from 11.2 to 50.7% and has varied widely with geographical location. Although no detailed information has been published on AE in domestic and wild animals in Turkey, the main definitive and intermediate hosts of E. multilocularis are assumed to be rodents and red foxes, respectively. However, there has been only a single published report of E. multilocularis in a wild animal (fox) in the northwest in 1965 by Merdivenci. The first human case of AE in Turkey was reported by Mutlu in 1939, and total cumulative reported case number is 202 between 1980 and 1998.