A comparative analysis of different molecular targets using PCR for diagnosis of old world leishmaniasis.

The different sensitivity values were obtained in each study conducted for the diagnosis of leishmaniasis with the polymerase chain reaction (PCR). However, a standardized PCR target for the diagnosis of leishmaniasis does not exist. The aim of the current study, the most ideal PCR target was determined for diagnosis of leishmaniasis. A total of 72 smear and 48 bone marrow samples were analyzed with six different molecular targets to determine their potential as a tool for the specific molecular diagnosis of leishmaniasis using PCR. The positivity-negativity value and the sensitivity-specificity of each PCR targets were calculated. The positivity value of PCR targets were sequenced in different levels in the diagnosis of leishmaniasis from highest to lowest in the order of kDNA-PCR > SSU rRNA-PCR > ITS2-PCR > ITS1-PCR > ME-PCR > HSP70-PCR. The sensitivities of PCR targets except ITS1-PCR, ME-PCR and HSP70-PCR were found to be 100% in cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) cases as compared to microscopic examination accepted as a gold standard. The sensitivities of ITS1-PCR, ME-PCR and HSP70-PCR were found 96.6%, 90.0% and 86.6%, respectively, in CL-cases. In addition, the sensitivities of ITS1-PCR, ME-PCR and HSP70-PCR were found 90.0%, 70.0% and 60.0%, respectively, in VL-cases. The kDNA genomic region was the most sensitive for routine diagnosis of leishmaniasis. ITS1-PCR restriction fragment length polymorphism, the alternative method for the identification of Old World Leishmania species, did not require culturing of the parasites.

Clinical manifestations and genetic variation of Leishmania infantum and Leishmania tropica in Southern Turkey.

L. infantum was isolated from cutaneous leishmaniasis (CL) skin lesions in patients having no signs and symptoms of visceral leishmaniasis (VL). Similarly, L. tropica had previously been isolated from patients with VL in the absence of cutaneous lesions. It was not certain how visceralization occurred. Smears (207) and bone marrow samples (135) were taken from CL and VL-suspected patients, respectively. Microscopic examination, ITS1-PCR, RFLP and DNA sequencing for all samples were analyzed. The microscopic examination of smears was found to be 61.3% (127/207) in CL-suspected cases and bone marrow samples were found to be positive 8.8% (12/135) in VL-suspected cases. L. tropica 48.6% (72/148), L. infantum 35.8% (53/148), L. major 15.6% (23/148) in CL, and L. infantum 56.3% (18/32), L. donovani 31.2% (10/32), L. tropica 12.5% (4/32) in VL were found with PCR-RFLP. In addition, the DNA sequencing revealed a genetic variation in L. infantum (variants 1-3) and L. tropica (variants 1-5). We assume that the increased disease occurrence may have resulted from geographical expansion of disease, changing patterns of international travel, population migrations, non-immune people into endemic regions of infected people into non-endemic regions. In this study, L. infantum (variant 3) only in CL-patients and L. tropica (variant 2) only in VL-patients were identified. We hypothesize that genetic variation might play a role in the causation of CL and VL in southern Turkey and the genetic variants may differ according to the geographical location among Leishmania strains.

Genotyping of Acanthamoeba T15: the environmental strain in Turkey.

BACKGROUND: 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 rRNA-DNA 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 emergence of Leishmania major and Leishmania donovani in southern Turkey.

BACKGROUND: In southern Turkey, Leishmania tropica and L. infantum are both the causative agents of cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL), respectively. However, L. major and L. donovani were known to exist after the influx of Syrian refugees. METHODS: Between the years of July 2003 and July 2013, a total of 167 smears and 113 bone marrow samples were taken from CL and VL-suspected cases, respectively. Samples were analysed through real-time PCR and ITS1 DNA sequencing. RESULTS: One hundred and seven 64% (107/167) smears and 56% (63/113) bone marrow samples were positive for leishmaniasis according to the real-time PCR. Three different Leishmania species were found in the 107 CL cases by real-time PCR: 42% (45/107) L. tropica, 36.5% (39/107) L. infantum and 21.5% (23/107) L. major. In addition, three different Leishmania species were identified in the 63 VL cases: 60.3% (38/63) L. infantum, 30.2% (19/63) L. donovani and 9.5% (6/63) L. tropica using real-time PCR. The results of real-time PCR were confirmed with Leishmania ITS1 DNA sequencing. CONCLUSIONS: This study revealed that in southern Turkey, L. major and L. donovani were the aetiological agents of CL and VL, respectively. It was assumed that emergence of L. major and L. donovani was due to influx of Syrian refugees, as well as the effects of global warming.

Oxidative stress in vivax malaria.

Malaria is still a leading cause of morbidity and mortality. The increase in lipid peroxidation reported in malaria infection and antioxidant status may be a useful marker of oxidative stress during malaria infection. The aim of this study was to investigate the role of antioxidant enzymes against toxic reactive oxygen species in patients infected with Plasmodium vivax and healthy controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were determined in 91 P. vivax patients and compared with 52 controls. Malondialdehyde levels, superoxide dismutase, and glutathione peroxidase activities were 8.07±2.29 nM/ml, 2.69±0.33 U/ml, and 49.6±3.2 U/g Hb in the patient group and 2.72±0.50 nM/ml, 3.71±0.47 U/ml, and 62.3±4.3 U/g Hb in the control group, respectively. Malondialdehyde levels were found statistically significant in patients with vivax malaria higher than in healthy controls (P<0.001). On the other hand, superoxide dismutase and glutathione peroxidase activities were found to be significantly lower in vivax malaria patients than in controls (P<0.05). There was an increase in oxidative stress in vivax malaria. The results suggested that antioxidant defense mechanisms may play an important role in the pathogenesis of P. vivax.

Evaluating the trace elements in preoperative and postoperative duration of hydatid surgery.

OBJECTIVE: To evaluate the preoperative and postoperative values of serum levels of zinc, magnesium, and copper in patients with cystic echinococcosis (CE). METHODS: This study was conducted on patients with CE between 2000-2005 at the Faculty of Medicine, University of Cukurova, Adana, Turkey. Serum levels of zinc, copper, and magnesium of 85 patients with CE were measured both before and one year after the operation. Patients with liver CE (85 patients, 48 males) and healthy adults (40 adults, 17 males) as the control group were enrolled in the study. RESULTS: When compared with the control group, the mean serum levels of zinc and magnesium were lower and the mean serum level of copper was higher in the preoperative period. It was found that as duration of symptoms increased, serum zinc and magnesium levels decreased and copper levels increased. The serum levels of these elements in postoperative and control patients after one year compared equally with the levels in the initial control. CONCLUSION: The probability of increased consumption of zinc and magnesium and secretion of copper by the parasite may lead to evaluating the follow up of hydatid surgery in addition to radiological and serological methods.

Supportive presumptive diagnosis of Plasmodium vivax malaria. Thrombocytopenia and red cell distribution width.

OBJECTIVE: To evaluate hemoglobin, leukocyte, platelet counts and red cell distribution width values during acute vivax malaria. METHODS: This study, which comprises 90 symptomatic vivax malaria patients compared with 52 healthy controls, investigated hemoglobin, leukocyte, platelet counts and red cell distribution width values during acute disease prior to the treatment in vivax malaria, from May 2002 to December 2004 in Adana, Cukurova region, located in the southern part of Turkey, along the Mediterranean coast. RESULTS: Mean values for hemoglobin, leukocyte and platelet counts in the vivax malaria group in our study were found to be significantly lower in comparison to the control group. Anemia and thrombocytopenia were also observed in the malaria group while not in the control group (p<0.05, p<0.0001). Mean red cell distribution width values were found to be significantly higher in the malaria group (p<0.0001). CONCLUSION: Our findings indicated that routinely used laboratory findings such as low hemoglobin, leukocyte or platelet counts and especially high red cell distribution width values could present a more supportive clue in the diagnosis of vivax malaria in endemic areas.

Serum malondialdehyde level in patients with cystic echinococcosis.

OBJECTIVE: To investigate the changes of serum malondialdehyde (MDA) level for the oxidative stress hypothesis in patients with cystic echinococcosis (CE). METHODS: The study was conducted on patients with CE before the surgical treatment. Anti-Echinococcus granulosus antibodies were determined by serological method. We assayed MDA activities measured of 57 subjects and matched in 45 healthy controls. This study took place in the Faculty of Medicine, University of Cukurova, Balcali Hospital in Adana, Turkey, between March 2004 and October 2005. RESULTS: The mean +/- SD of MDA levels of patients with CE was 6.70 +/- 1.66 and healthy controls was 2.53 +/- 0.43. The difference between MDA levels of patients and controls was statistically significant (p<0.001). CONCLUSION: The high infection/control ratio of MDA concentration and the significant correlation strongly indicate the occurrence of oxidative stress and lipid peroxidation as a mechanism of tissue damage in cases of CE. Our study highlights the usefulness of serum markers to investigate complex pathological situations, including distinct forms of chronic liver inflammation associated with CE.