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Evaluation of the transmission mode of B. hominis by using PCR method

PubMedID: 20544220Blastocystis hominis is a common intestinal parasite observed in fecal examination. On the other hand, the transmission of this parasite is certainly unknown. The transmission of B. hominis can be realized by animal contact and the contamination by water and food with excreted cysts from the reservoir hosts. B. hominis isolated from 25 humans, their pets, and tap water was identified by polymerase chain reaction using sequenced tag site primers in this study. B. hominis isolates obtained from humans and pets were identified as subtype1, subtype2, and subtype3 while B. hominis isolates obtained from tap water were also identified as subtype1. The B. hominis isolates obtained from humans in this study were defined as the same as the subtypes of the B. hominis isolates obtained from the pets, of which these people keep at their homes, and the tap water. These findings reveal that the source of B. hominis infection could be pets and tap water. © 2010 Springer-Verlag

Subtype analysis of Blastocystis isolates using SSU rRNA-DNA sequencing in rural and urban population in southern Turkey

PubMedID: 27725159Blastocystis is a common and emerging parasite often seen in many studies conducted in urban population, with scanty reports on rural communities. However, little is known about the public health significance of Blastocystis infection. A total of 28 Blastocystis isolates from 17 (17/28, 60.71%) patients living in rural area and 11 (11/28, 39.29%) patients living in urban area were screened with seven kinds of sequenced-tagged site primers for identification of subtype. PCR products were sequenced with same combination of primers using the BigDye Terminator V 3.1 cycle sequencing kit, as per the manufacturers’ protocol on the 3730 DNA analyzer (Applied Biosystems, Carlsbad CA, USA). The cross-comparison of the Blastocystis sequences of samples were determined by the neighbor-joining method based on a distance matrix between sequence pairs to generate dendograms. The following subtypes were identified; subtype 1 (10/28, 35.7%), subtype 3 (7/28, 25.0%), subtype 2 (5/28, 17.8%), subtype 4 (3/28, 10.7%), subtype 5 (1/28, 3.6%), subtype 6 (1/28, 3.6%), and subtype 7 (1/28, 3.6%) in all DNA samples. The comparison of Blastocystis subtypes distribution among the patients from rural and urban area revealed subtype 5 (1/17, 5.9%), subtype 6 (1/17, 5.9%) and subtype 7 (1/17, 5.9%) from patients of rural area but not any of these subtypes in patients living urban area. This study is the first large-scale study to examine the occurrence of Blastocystis in Turkey to shed lights on the cosmopolitan distribution of Blastocystis subtypes in southern part of Turkey. Subtype 5, subtype 6 and subtype 7 were determined in only rural area. The findings of this study suggest that Blastocystis is transmitted from animal to human and possess a zoonotic potential. © 2016 Elsevier Inc

Investigation of Cryptosporidium spp. antigen by ELISA method in stool specimens obtained from patients with diarrhea

PubMedID: 20938687Cryptosporidium spp. is an important parasitic protozoan causing diarrhea in developing and developed countries. The agent causes severe life-threatening diarrhea especially in immunocompromised hosts. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive and time-consuming. Thus, we aimed to evaluate the usefulness of a copro-antigen enzyme-linked immunosorbent assay (ELISA) test in detecting Cryptosporidium spp. from fecal specimens. For this aim, microscopy and specific antigen detection methods were compared to determine Cryptosporidium spp. In addition, specific antigen by ELISA method in stool was investigated in order to find out whether or not it contributes to the diagnosis of Cryptosporidium spp. One hundred and fifty-four stool specimens taken from patients whose ages ranged from 0 to 86 with diarrhea applied to Department of Parasitology, Balcali Hospital of Cukurova University in Adana, Turkey were used. All samples were examined for Cryptosporidium spp. antigen by ELISA and oocysts via gold standard modified acid-fast staining, between October 2008 and July 2009. Eight (5.19%) specimens were found to be positive by modified acid-fast staining method and 37 (24.03%) specimens by copro-antigen ELISA method were found to be positive. The sensitivity and specificity for copro-antigen ELISA were 100% and 80.1%, respectively. The results of copro-antigen ELISA indicate that the simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis and may be useful for large-scale epidemiological studies of cryptosporidiosis. © 2011 Springer-Verlag.TF.2008Acknowledgements This study was supported by the Cukurova University Research Grant TF.2008.YL.

Comparison of clinical samples and methods in chronic cutaneous leishmaniasis

PubMedID: 25223940This study aimed at finding out the most effective clinical samples and methods in chronic cutaneous leishmaniasis (CCL). Smear, aspiration fluid, and filter paper samples were taken from 104 skin lesions of suspected cases with CCL, and they were compared using microscopic examination, culture, and molecular methods. We characterized four different forms of CCL and identified the causative agents in CCL forms using high-resolution melting curve real-time polymerase chain reaction assay. We observed that smear was detected to be the most sensitive (63.5%) among clinical samples, and real-time polymerase chain reaction method was the most sensitive (96.8%) among the methods used in diagnosis of CCL. We identified 68.8% Leishmania tropica and 31.2% L. infantum in papular lesions, 69.2% L. infantum and 30.8% L. tropica in nodular lesions, 57.9% L. tropica and 42.1% L. major in ulcerating plaque lesions, and 55.5% L. tropica and 44.5% L. major in noduloulcerative lesions in CCL patients. Copyright © 2014 by The American Society of Tropical Medicine and Hygiene

Genotyping of Acanthamoeba T15: The environmental strain in Turkey

PubMedID: 25424836Background: Environmental sources are potential sources for the transmission of Acanthamoeba in humans and other mammals. Methods: A total of 50 water samples from hot springs and swimming pools, and 50 soil samples were taken from Adana, Afyon, Kutahya, Mersin and Nigde provinces in Turkey. Samples were analysed using 18S rRNADNA sequencing. Results: Acanthamoeba griffini (T3), Acanthamoeba castellanii (T4) and Acanthamoeba jacobsi (T15) were found in water samples. Acanthamoeba griffini (T3) and Acanthamoeba castellanii (T4) were detected in soil samples. Conclusions: In Turkey, this was the first time that Acanthamoeba jacobsi (T15) was detected in water samples. © The Author 2014.TF.2011Funding: Funding was provided by the Cukurova University Research Grant [TF.2011.YL.05]

Detection of Plasmodium vivax by nested pcr and real-time PCR

PubMedID: 20585524Malaria is endemic in the Cukurova region while it is sporadic in other regions of Turkey. Therefore, the laboratory and clinical diagnosis of malaria is important for the treatment of malaria. In this study, 92 blood samples that were taken from the suspected malaria patients for routine diagnosis in a period of 10 years between 1999 and 2009 were analyzed. All of these blood samples were examined by microscopic examinations using Giemsa-stained thick blood films, nested PCR, and real-time PCR. The sensitivity-specificity and positive-negative predictive values for these diagnostic tests were then calculated. It was found that the positive predictive values of microscopic examination of thick blood films, nested PCR, and real-time PCR were 47.8%, 56.5%, and 60.9% for malaria, respectively. The real-time PCR was found to have a specificity of 75% and sensitivity of 100%, while specificity and sensitivity of nested PCR was found 81.2% and 97.7% according to the microscopic examination of thick blood films, respectively

Intestinal parasites in patients with hemopoietic neoplasia

Purpose: The aim of this study is to determine the incidence of intestinal parasites in patients with leukemia and lymphoma. For this aim 100 stool samples taken from patients were examined and compared with samples taken from healthy controls. The ages were between 15 and 80. Methods: Nativ-Lugol and Formalin-ether centrifugation methods were used in stool samples. Modified Ziehl-Neelsen and Sheather's sugar flotation methods were used to detect the Cryptosporidium species. Results: Parasite incidences were 15% and 10% in study and control groups respectively. Cryptosporidium species were not found both in study and control groups. Conclusion: As a result, parasite incidence was not found to be increased in patients who were immunosupressed as compared to healthy controls

Feconomics®; a new and more convenient method, the routine diagnosis of intestinal parasiticinfections

PubMedID: 24781020Direct wet mount examination and concentration are the most commonly used methods for detecting intestinal parasites from fecal samples. Concentration methods are used when there are fewer protozoan cyst, coccidian oocyst, microsporidial spore, helminth egg, and larvae in the fecal samples. Early detection of the causative intestinal parasites plays a significant role in implementing timely and correct treatment, which relieves the patients' symptoms and also prevents recurrences. Formalin-ethyl acetate concentration (FEAC) is believed to be a gold standard method to detect most intestinal parasites. Thus, in this study, we evaluated the diagnostic value of Feconomics® [manufactured by Salubris Inc, Boston, USA. Patent application number (TR): 2010/07549] which is a simple, new, and rapid fecal concentration method for the detection of the intestinal parasites in human beings. We also compared the FEAC with Feconomics® and direct wet mount examination. A total of 918 fecal samples were collected from the patients suspected to have intestinal parasitic infection. Samples were examined with the direct wet mount, FEAC, and Feconomics® methods. Different parasite species 15.9 % (146/918) with Feconomics®, 13.3 % (122/918) with FEAC, and 9.8 % (90/918) with direct wet mount examination, Feconomics®>FEAC>direct wet mount examinations were detected. They were statistically compared considering FEAC as the gold standard for parasitological diagnosis; the sensitivity and specificity of Feconomics® were calculated as 96 and 97 %, respectively. Blastocystis hominis was found to be the most common parasite, followed by Giardia lamblia with direct wet mount examination, FEAC, and Feconomics® methods. Feconomics® proved to be better than not only FEAC in concentrating parasite egg and cyst forms as well as in maintaining characteristic morphology but it is also better in direct wet mount examination. Feconomics® eliminates the need for centrifugation by using absorbent beads that help the homogenization and concentration of the sample. Feconomics® in this study was considerably better than FEAC in detecting the trophozoites of Giardia lamblia. We suggest that Feconomics® be used for the routine diagnosis of intestinal parasitic infection in rural areas of developing countries due to the fact that a centrifuge is not required and it eliminates large stool particles. © 2014 Springer-Verlag

Identification of Blastocystis hominis isolates from asymptomatic and symptomatic patients by PCR

PubMedID: 19685075Despite years of study, the pathogenic role of Blastocytis hominis is still controversial. Genotypic differences between the asymptomatic and symptomatic isolates should assist in determining the pathogenicity of Blastocystis. In this study, we genotyped 32 Blastocystis isolates obtained from 12 asymptomatic healthy individuals and 20 symptomatic patients pain by polymerase chain reaction using known seven kinds of sequence tagged site primers in this study. When we compared genotype of Blastocystis isolates between the symptomatic and asymptomatic patient group, we found that subtype3 is the most dominant genotype in asymptomatic individual (9/12) and subtype1 determined all of symptomatic patients (20/20). © 2009 Springer-Verlag.TF.2007Acknowledgements This study was supported by the Cukurova University Research Grant TF.2007.BAP.26