The importance of Real-Time Polymerase Chain Reaction method in diagnosis of intestinal parasites in cases with diarrhea.

Diarrheal diseases cause significant morbidity in developing countries, and have a potential to cause severe complications. Stool samples collected from 272 patients with diarrhea in Adana, Turkey were screened using different laboratory methods such as the wet mount examination (WME), modified trichrome stain (MTS), modified acid-fast (MAF) stain, enzyme linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (realtime PCR) for intestinal parasites. Intestinal parasites were detected in 25.4% (69/272) of the patients. Of the parasites detected, Cryptosporidium spp. was the commonest 10.3% (28/272) followed by Blastocystis spp., 5.9% (16/272), Entamoeba dispar 2.6% (7/272), Giardia lamblia 2.6% (7/272), Entamoeba coli 1.8% (5/272), Entamoeba histolytica 0.7% (2/272), Isospora belli 0.7% (2/272), Trichuris trichiura 0.4% (1/272) and Strongyloides stercoralis 0.4% (1/272). Our study shows that there are differences in results obtained using different laboratory methods; 22.1% (60/272) with real time-PCR, 10.3% (28/272) with ELISA, 9.9% (27/272) with MTS, 6.6% (18/272) with WME, and 5.9% (16/272) with MAF stain. The differences in the level of positivity using different laboratory methods were sequenced in the diagnosis of intestinal parasites ranging from highest to lowest, as real-time PCR>ELISA>MTS> WME>MAF stain. Molecular techniques are recommended as diagnosis of choice especially in patients suffering from chronic diarrhea.

Molecular Identification and DNA Sequencing of Trichomonas vaginalis Strains from Agean region of Turkey.

The aim of this study is to compare the presence of Trichomonas vaginalis (T. vaginalis) in symptomatic and asymptomatic women through microscopic examination, culture in Trypticase-Yeast Maltose (TYM) medium and PCR methods. In addition, T. vaginalis strains were analysed for genotyping with 18S rRNA-DNA and phylogenetic analysis. Axenized strains of T. vaginalis isolated from urine culture samples taken from symptomatic and asymptomatic women with clinical signs. Molecular characterization of the isolated strains of T. vaginalis was performed by using PCR. To evaluate molecular diagnosis and genotypic identification of T. vaginalis strains, 14 samples were analysed. Of the 14 samples, T. vaginalis was positive in 14 samples by microscopy, 6 in culture( TYM medium) and 14 by PCR, respectively. Although the sample size is very small, PCR was shown to be high sensitivity and specificity, and seems to be a promising diagnostic tool. 18S rRNA-DNA PCR results also confirmed with real time PCR method. In conclusion, it is considered that two strains of T. vaginalis isolated from samples, 5-TV1G and 13-TV1G, are subtypes of T. vaginalis as a result of 18S rRNA-DNA sequencing analysis. To best of our knowledge this is the first analysis of phylogenetic positions on T. vaginalis from Turkey.

Molecular Identification and DNA Sequencing of Trichomonas vaginalis Strains from Agean region of Turkey

The aim of this study is to compare the presence of Trichomonas vaginalis (T. vaginalis) in symptomatic and asymptomatic women through microscopic examination, culture in Trypticase-Yeast Maltose (TYM) medium and PCR methods. In addition, T. vaginalis strains were analysed for genotyping with 18S rRNA-DNA and phylogenetic analysis. Axenized strains of T. vaginalis isolated from urine culture samples taken from symptomatic and asymptomatic women with clinical signs. Molecular characterization of the isolated strains of T. vaginalis was performed by using PCR. To evaluate molecular diagnosis and genotypic identification of T. vaginalis strains, 14 samples were analysed. Of the 14 samples, T. vaginalis was positive in 14 samples by microscopy, 6 in culture( TYM medium) and 14 by PCR, respectively. Although the sample size is very small, PCR was shown to be high sensitivity and specificity, and seems to be a promising diagnostic tool. 18S rRNA-DNA PCR results also confirmed with real time PCR method. In conclusion, it is considered that two strains of T. vaginalis isolated from samples, 5-TV1G and 13-TV1G, are subtypes of T. vaginalis as a result of 18S rRNA-DNA sequencing analysis. To best of our knowledge this is the first analysis of phylogenetic positions on T. vaginalis from Turkey.

Entomological Survey for Sand Fly Fauna in Imamoglu Province (Cutaneous Leishmaniasis Endemic Region) of Adana, Turkey.

Leishmaniasis, presenting in two clinical forms, cutaneous and visceral in Turkey, is widespread in most of the countries in the Mediterranean Basin. An average of 10 to 13% of cases are reported from Adana every year. This paper presents the results of an entomological survey in an endemic focus of cutaneous leishmaniasis in Imamoglu province of Adana in Turkey.We collected 654 sand fly specimen using 100 light traps in 20 nights in August 2013 and July 2014. Several keys and previous drawings were used in the identification of the species. In total, six Phlebotomus species were identified; Phlebotomus tobbi (50.3%), Phlebotomus papatasi (34%), Phlebotomus perfiliewi (2.9%), Phlebotomus sergenti (0.4%), Phlebotomus neglectus/syriacus (0.8%), and Sergentomyia spp. (11.5%). In addition, the female/male rate was found to be 0.84.Collected sand flies (44 pools of 1-10 individuals) were analyzed microscopically, and no promastigotes were found in the midgut specimens. Using a genus specific real time-polymerase chain reaction (PCR) assay for simultaneous detection of Old Word Leishmania. We detected only 3 of the 44 pools with Leishmania by genus-specific real-time PCR assay.P. tobbi was found to be dominant species in spite of the differences in sand fly fauna composition. To the best of our knowledge, this is the first study for sand fly fauna including 15 villages where endemic focus of cutaneous leishmaniasis in Imamoglu, Adana Province, Turkey.

Prevalence of Dirofilaria immitis in dogs from Hatay province, Turkey.

The present study was conducted to determine the prevalence of canine dirofilariosis in Hatay province, south of Turkey. A total of 269 blood samples were collected from owned dogs in this multi-centre survey between March and July 2006. Blood samples were examined by modified Knott and enzyme-linked immunosorbent assay (ELISA) techniques to detect circulating microfilariae and antigens of Dirofilaria immitis, respectively. Seventy out of 269 dogs (26.0%) were positive for D. immitis. However, 61.4% of positive dogs had occult infection. The prevalence of canine dirofilariosis was heterogeneous in Hatay province, with higher values in shoreline (33.0%) and riverside (30.9%) areas followed by the lowland (25.5%) or mountainous (15.2%) areas. No statistically significant differences were observed in relation to sex (females, 33.3%; males, 24.4%, P>0.05). When evaluating the prevalence of D. immitis by age, the highest prevalence was observed in dogs older than 4 years of age. Large breeds (29.6%) and the dogs living outdoors (30.2%) showed a higher prevalence in comparison to small breeds (21.4%) and the dogs living indoors (10.5%), respectively. In conclusion, according to the results of this study canine dirofilariosis had a high prevalence in Hatay province. Therefore, prophylaxis against heartworm is advisable to decrease the incidence of canine dirofilariosis.

Alveolar hydatid disease: imaging findings and surgical approach.

PURPOSE: We report six cases of Alveolar Hydatid Disease (five in the liver and one in the gastrosplenic ligament invading the spleen) in which curative resection of the liver (five cases) and splenectomy (one case) were performed. MATERIAL AND METHODS: The records of the six patients with AHD were retrospectively evaluated. Demographics of the patients, symptoms, laboratory findings, including serology were recorded. Imaging studies determined the extent of the disease preoperatively. Classification of the lesions was done according to the PNM (P = parasitic mass in the liver, N = involvement of neighbouring organs, and M = metastasis) staging system designed by the World Health Organization. All the surgical procedures were performed as complete resections, where negative margins were approved by frozen sections. Chemotherapy with albendazole (10 mg/kg/day) was continued postoperatively for two years in five of the six patients who were alive. RESULTS: All of the cases were from East Anatolia of Turkey, which is an endemic region. The mean age was 39.6 years (15-54 years). Major complications occurred post-operatively in all patients, possibly due to the extensive resection. No recurrence was seen during the 5 year follow-up of two cases and 2 year follow-up of three cases. CONCLUSION: The treatment of Alveolar Hydatid Disease is curative radical resection. Thus, pre-operative imaging studies to determine the extent and stage of the disease are of crucial importance.

Surgery in Fasciola hepatica pancreatitis: report of a case and review of literature.

Fasciola hepatica is a trematode rarely causing disease in humans. In symptomatic cases, while various pathologies such as damage to liver parenchyma, acute cholecystitis, and obstructive jaundice can be seen, the development of pancreatitis is rarely mentioned in the literature. The treatment of the disease is medical. In cases where no definite diagnosis can be made or in incidental cases where common bile duct exploration is being done, F. hepatica can be detected accidentally during operation. No consensus has yet been reached on the surgical procedure to be applied in this condition. We report on our case due to the rare occurrence of pancreatitis as a complication. In surgical cases, external drainage of the bile is both crucial in observing the response to the treatment, and also should be accepted as part of the treatment.

Importance of the detection of amoebic antigens in stool samples for the diagnosis of Entamoeba histolytica infection, among children in southern Turkey.

Entamoeba histolytica is the predominant causative agent of human amoebiasis, a significant and common diarrhoeal disease among children of developing countries. Diagnosis of this illness by the microscopical detection of the parasites in stool samples is insensitive and often incapable of differentiating the pathogenic Entamoeba histolytica from the commensal parasite E. dispar. In this study, the results of testing stool samples in an ELISA, based on a monoclonal antibody, that detects E. histolytica-specific galactose adhesin were compared against the results of the microscopical examination of the same samples. The samples investigated came from 131 children (aged<15 years) with diarrhoea, who lived in the provinces of Adana and Mersin, in southern Turkey. Overall 22 cases of E. histolytica infection, including eight that appeared negative by microscopy, were detected using the ELISA. The 16 patients considered positive for E. histolytica/E. dispar by microscopy included two who were ELISA-negative. With the ELISA results used as the 'gold standard', and assuming that all the E. histolytica/E. dispar cysts seen by microscopy were E. histolytica, microscopy had a specificity of 98.2%, a positive predictive value of 87.5% and a negative predictive value of 93.1% but a sensitivity of only 63.6%. Compared with microscopy, culture and PCR-based assays, the antigen-detection ELISA appears to be easier, faster and probably more cost-effective, with high sensitivity, specificity and predictive values.

Serum copper, zinc and magnesium levels in children with enterobiosis.

Levels of serum copper (Cu), zinc (Zn) and magnesium (Mg) were determined in the sera of 250 children aged between 6 and 13. Of these children, 180 were infected only with Enterobius vermicularis. The remaining 70 children were without parasitic or bacterial infection and made up the control group. The cellophane tape method was used to detect E.vermicularis infection. The levels of Cu, Zn and Mg in the serum samples were measured with the Perkin- Elmer 2380 Atomic Absorption Spectrophotometer. Evaluation by the student-t test showed that the means of the Cu, Zn and Mg in the serum were significantly lower in the infected group than in the control group. Thus, in this study, we found that E. vermicularis adversely affects the level of elements such as Cu, Zn, and Mg in serum.