Seroprevalence of toxoplasmosis among individuals with intellectual disability in Hormozgan Province, southern Iran.

BACKGROUND: People with mental defects are more likely to get the infection due to their low levels of health care and personal hygiene. The current study aimed to determine the seroprevalence of Toxoplasma infection among individuals with intellectual disabilities in Hormozgan province, southern Iran. METHODS: The study population was 117 individuals with intellectual disabilities. Venous blood (3 mL) was taken from each subject. A commercial ELISA kit was used to determine anti-Toxoplasma IgG antibodies. RESULTS: Of 117 recruited subjects, 55 (47.0%) were men and 62 (53.0%) were women. The mean age of participants was 27.6 (±12.31) years. Out of 117 studied subjects, 76 had severe and 41 had profound intellectual disabilities. Anti-Toxoplasma IgG antibodies were detected in the sera of 35 out of 117 (29.9%) individuals. Seropositivity to toxoplasmosis was significantly higher in severe than in individuals with profound intellectual disabilities (P < 0.05). There was no statistically significant association between Toxoplasma infection and age, sex and duration of residency in the rehabilitation centre. CONCLUSIONS: The findings of this study indicate that the prevalence of Toxoplasma in people with intellectual disabilities is not much different from other groups of the community.

Stereological analysis of liver, spleen and bone of Leishmania infantum-experimentally infected hamsters.

Leishmaniasis is remaining as one of the important health problems of many countries around the world. The histopathology of the disease and the effects of the parasite on various tissues have not yet been fully elucidated. The current study aimed to evaluate the stereological features of the liver, spleen, and bone of hamsters infected with Leishmania infantum. In this experimental study, the L. infantum parasite was mass cultivated in a culture medium. Then, 15 golden hamsters were selected, of which 5 animals were considered as controls and another 10 animals were injected intravenously, with 1 × 108 promastigotes of L. infantum. Four months later, the hamsters were euthanized and impression smears were prepared from the liver and spleen. Moreover, pathology slides were prepared from the spleen, liver, and femur. The orientated method was used to obtain isotropic uniform random (IUR) sections. For stereological evaluation, the tissues were fixed with formalin buffer, and sections (4 and 25 µm thick) were prepared and stained with Heidenhain's AZAN trichrome and hematoxylin-eosin, respectively. The tissue samples were examined by stereological methods and all changes in the samples of the infected hamsters were compared with the control group. The number of hepatocyte and their nuclei volumes were significantly decreased in the Leishmania-infected group, compared to the control group. The number of Kupffer cells and their volume in the liver of the Leishmania-infected group was higher than that of the control group, and the differences were statistically significant. The volume of trabeculae and central arteries in the spleen of the Leishmania-infected group was lower than that of the control group and the number of lymphocytes and macrophages in the spleen of the Leishmania-infected group was increased compared to the control group. The trabecular volume and the number of osteoblasts and osteoclasts of the femur in Leishmania-infected animals decreased, whereas the volume of bone marrow was significantly raised. Leishmaniasis leads to changes in tissue structure and their function in the host by the involvement of various organs of the immune system including the liver, spleen, and bone. Understanding these changes are important in identifying the effective mechanisms of the parasite and host interaction.

Toxoplasmosis in Nomadic Communities: A Seroepidemiological Study in Southwestern Iran.

BACKGROUND: Toxoplasma gondii is the agent of a zoonotic protozoan infection with worldwide distribution. The rates of infection in different areas are depending on climatic conditions and also social and cultural habits of the people. This cross-sectional study aimed to find out the seroprevalence of toxoplasmosis in nomadic people in Boyer-Ahmad County, in the southwest of Iran. METHODS: Blood samples (5 mL) were collected from 1005 individuals of nomadic tribes of Boyer-Ahmad County in Kohgiluyeh and Boyer-Ahmad Province in 2016. A predesigned questionnaire, containing basic epidemiological information was filled out for each subject during the sampling. Sera were evaluated for anti-T. gondii antibodies (both IgG and IgM), using a commercial ELISA kit. Collected data were analyzed by SPSS, using descriptive statistics and Chi-square test. RESULTS: Out of the 1005 studied subjects, 227 (22.6%) were male and 778 (77.4%) were female. The mean age of participants was 41 years (SD: ±17). The most represented (24.4%) age class was 21-30 years and the least (1.9%) 1-10 years. Anti-T. gondii antibodies were detected in 174 (17.3%) of the cases. No statistically significant associations were found between seropositivity to T. gondii and area of residence, age, gender and educational levels of the participants. CONCLUSIONS: Findings of this study show that the seroprevalence of toxoplasmosis among the nomads of Kohgiluyeh and Boyer-Ahmad province is alike to other communities in our country. Further study is required to elucidate the genotype of T. gondii in this community.

High frequency of subclinical Leishmania infection among HIV-infected patients living in the endemic areas of visceral leishmaniasis in Fars province, southern Iran.

Visceral leishmaniasis (VL) is a major health concern in patients with HIV infection in endemic areas of VL. In these areas, a substantial number of infected individuals are asymptomatic and the risk of acute VL infection in HIV/VL co-infected cases is high. The current study aimed to determine the prevalence of asymptomatic VL infection among HIV-infected patients in Fars province, southern Iran. Subjects of the study were 251 HIV-confirmed patients who all were clinically asymptomatic for leishmaniasis. Blood samples were obtained from each participant. Anti-Leishmania antibodies were detected in the sera using ELISA. DNA was extracted from the buffy coat of each subject and PCR amplified, targeting an ITS-2 gene of Leishmania. PCR products were purified from the gel and were sequenced. Overall, 19 out of 251 (7.6%) HIV-infected patients were found to be infected with Leishmania, using serological or molecular methods. Anti-Leishmania antibodies were detected in 13 (5.2%) patients and leishmanial DNA in 8 (3.2%) of the patients. The sequence analysis of DNA-positive cases revealed the species of the parasite as L. infantum. The high prevalence of VL among the patients with HIV is a serious challenge which demands further attention to improve the prophylaxis and treatment measurements of VL/HIV co-infection and thereby promoting the life expectancy and quality of life of these patients.

Genetic diversity of Fasciola spp. isolates from northern part of Iran: comparison with southwestern isolates.

Animal and human fascioliasis is a health and economic problem in few of tropical and subtropical areas of the world, including Iran. The present study aimed to determine the genotype diversity of Fasciola isolates in different hosts from Gilan province, northern Iran, and compare it with those isolates from southwestern Iran. Forty-eight adult Fasciola spp. were collected from cattle, sheep, and goats from slaughterhouse in Talesh, north of Iran. DNA was extracted from each fluke and PCR-RFLP was used to find out the species of the isolates. The ribosomal ITS1 and ITS2, and mitochondrial genes of NDI and COI from individual Fasciola isolates of each host were PCR-amplified and the PCR products were sequenced. Genetic variation within and between the isolates was evaluated by comparing the sequences of ribosomal and mitochondrial genes. For analysis of phylogenetic diversity of the flukes, phylogenetic trees were constructed, using ITS1, ITS2, NDI, and COI sequences of the isolates. Based on PCR-RFLP profile, 5 (22.7%) of the total of sheep isolates and 18 (90%) of cattle isolates were identified as F. gigantica and other remaining samples from sheep, cattle and goats were identified as F. hepatica. Based on ITS1 and ITS2 sequences, six and seven nucleotide polymorphism were respectively noted in the isolates. On the other hand, CO1 region sequences showed considerable variation, which laid Talesh (north) isolates in a separate cluster. Findings of the study showed that the sequences of CO1 isolates from north and southwest have substantial differences mainly in CO1 region.

Detection of Fasciola hepatica and Fasciola gigantica common and uncommon antigens, using rabbit hyper immune serum raised against their excretory-secretory and somatic antigens.

Fasciolosis is an important neglected helminth disease caused by two liver flukes, Fasciola hepatica and Fasciola gigantica. The two species of Fasciola are usually different in their morphological and molecular features. They have also common and uncommon antigens in both their somatic and excretory secretory metabolites. In this study, we compared somatic and excretory-secretory (ES) antigens of F. hepatica and F. gigantica, by using rabbit hyper immune serum raised against these antigens. Adult worms were collected from bile ducts of infected animals and species of the fluke was confirmed by RFLP-PCR. ES and somatic antigens of both species were prepared. Rabbits were subcutaneously immunized with either ES or somatic antigens to produce antibodies against these antigens. SDS-PAGE pattern of F. hepatica and F. gigantica somatic antigens was similar and both of them revealed 30 protein bands, ranging from 18 to 180 kDa. In contrast, SDS-PAGE pattern of ES antigen of the two species was different. While protein bands with molecular weight of 18, 27, 29, 48, and 62 kDa were common in both species, bands of 19, 45, 55 and 58 kDa were only noticed in F. hepatica ES antigen. Rabbit polyclonal antibodies, raised against F. hepatica and F. gigantica ES antigen, reacted with main five protein bands, 25, 27, 29, 62 and 67 kDa and polyclonal antibodies raised against somatic antigens of both species reacted with three protein bands, 25, 27 and 72 kDa. Thus, the 25, 27 and 29 kDa protein bands may serve as immunodominant antigens, which might be considered for serodiagnosis of fasciolosis. Moreover, bands of 62 and 67 kDa in ES antigen and 72 kDa in somatic antigens of both species were immunodominant and might be suitable candidate for development of serological assays for diagnosis of fasciolosis.

Macracanthorhynchus hirudinaceus: the most common helminthic infection of wild boars in southwestern Iran.

A pathological and epidemiological study on Macracanthorhynchus hirudinaceus infection on 25 wild boars Sus scrofa in southwestern Iran was done. Overall 24 helminthes (18 female and 6 male) were collected from the intestine of 13 infected wild boars. Thus, the prevalence of acanthocephalans among the 25 examined boars was 52 %. Pathological evaluation revealed damages caused by the acanthocephalans penetrated in the small intestine tissue. Helminthes caused severe necrosis with ulcer and inflammation in tunica sub mucosa layer. High prevalence of this helminthic infection may cause severe hazard for farmers, residents and also wild life animals.

Comparative study on isoenzyme patterns of Fasciola hepatica and Fasciola gigantica.

Differences in the number of isoenzymes and their electrophoretic mobility are very valuable tools for characterization of closely related species of a certain parasite. This study aimed to compare the isoenzymatic patterns of Fasciola hepatica and Fasciola gigantica. Adult F. hepatica and F. gigantica were collected from infected livers of sheep and cattle and their species was determined by molecular method. Enzymes were extracted from the adult worms and subjected to electrophoresis through a polyacrylamide gel. The activities of nine enzymatic systems; including superoxide dismutase (SOD), glucose phosphate isomerase (GPI), glucose 6-phosphate dehydrogenase (G6PD), malate dehydrogenase (MDH), malic enzyme (ME), nucleoside hydrolyse 1 (NH1), phosphoglucomutase (PGM), isocitrate dehydrogenase (ICD), and 6-phosphogluconate dehydrogenase (6PGD) were evaluated from both Fasciola species. The enzymatic profile obtained for SOD system for F. hepatica were two-banded pattern whereas in F. gigantica it was a four-banded pattern. NH1 revealed one band in F. hepatica with relative mobility of 0.05 and two bands in F. gigantica with relative mobility of 0.05 and 0.66. In ICD, GPI and G6PD enzyme systems, both Fasciola species revealed one band but with different relative mobility. Isoenzymatic profile of MDH, 6PGD, PGM and ME were the same in both species. Findings of this study revealed that F. hepatica and F. gigantica have entirely different isoenzyme patterns in the enzymes of ICD, G6PD, GPI, PGM and SOD. These enzyme systems may be used for differentiation of these two species of Fasciola.

Seroprevalence of Leishmania infection among the healthy blood donors in kala-azar endemic areas of Iran.

Visceral leishmaniasis (VL) is a serious health problem in Iran. The disease is mainly transmitted by sand fly bites, but its transmission through transfusion in endemic areas may also occur. The current study aimed to determine the prevalence of Leishmania infection among blood donors in VL-endemic areas in south of Iran. A total of 2003 healthy blood donors from blood service centers in five VL-endemic districts in Fars province, southern Iran, were enrolled in the study. The blood samples were assessed for antibodies against Leishmania infantum by direct agglutination test (DAT). Seropositive subjects were tested for the presence of L. infantum DNA in their buffy coat by the molecular method. Socio-demographic features of the subjects were also documented during sample collecting. The mean age of participants was 36.3 (SD = 10.7 years). Male constituted 94.7 % of the subjects while only 5.3 % of donors were female. Twenty-eight blood donors (1.4 %) were positive for Leishmania infection by DAT. Only one of these seropositive donors was positive for Leishmania infection by polymerase chain reaction. A significant correlation was found between age, the place of residence and seropositivity to Leishmania (P < 0.05). Findings of this study revealed that the prevalence of Leishmania infection among blood donors in transfusion centers in the VL-endemic areas in Iran is relatively high. These asymptomatic blood donors may constitute a risk of transmitting of VL to susceptible recipients.

Epidemiological features of visceral leishmaniasis in fars province, southern iran.

BACKGROUND: To describe the epidemiological features of pediatric visceral leishmaniasis in southern Iran. METHODS: This retrospective study was carried out using local hospital records of VL patients from 2001 through 2009. Data such as age, gender, place of residence, clinical signs and symptoms, treatment, history of recurrence were recorded. The collected data were statistically analyzed using SPSS software. RESULTS: A total of 260 cases of VL have been recorded during 2001 to 2009 in south of Iran, based on hospital records. Mean age of patients was 3.5 years with the highest prevalence in 2 years old patients. The diseases have been more common in males (60%). The main clinical signs and symptoms of the patients were fever (96.2%), hepatosplenomegaly (68.8%) and abdominal protrusion (71.9%). Most of cases were from Kazeroun County (17.5%) in Fars Province followed by Borazjan in Boushehr Province. Bone marrow aspirations have been performed in 178 of cases and Leishmania amastigotes were detected in only 50 (28.1%) cases. Glucantime has been the first drug treatment while 19.3% of cases have been treated with amphotericin B. Mortality rate was 6.2% and children under 1 year old have been the main victim of the disease. Relapse has been noted in 7.3% of patients. CONCLUSION: VL is still endemic in South of Iran, especially in Fars Province with a noticeable mortality rate. Moreover, cases of the diseases are reporting from neighboring provinces and this might be due to the spreading of the diseases to the adjacent provinces.

Seroprevalence of human fasciolosis in a new-emerging focus of fasciolosis in yasuj district, southwest of iran.

BACKGROUND: Fasciolosis is an important health and veterinary problem in Iran. The epidemiological pattern of disease has been changed markedly in recent years and there are regions that have potent capacity to be new focus of the disease. One of these areas is Yasuj district in southwest of Iran where animal fasciolosis has been quite common. The current study was conducted to determine the seroprevalence of human fasciolosis in this area and to reveal the epidemiological factors associated with the spreading of the disease in this region. METHODS: One thousand blood samples were randomly collected from five villages in Yasuj district. ELISA, using Fasciola somatic antigen (SA), was carried out to detect anti Fasciola antibodies in the collected sera. RESULTS: Anti-Fasciola antibodies were detected in serum of 18(1.86%) individuals by ELISA. Out of 18 seropositive people, 9 (0.9) were female and 9 (0.9%) were male. Most of people (99.8%) had a history of consuming wild freshwater plants mainly Nasturtium microphyllum (local name Bakaloo) and/or Mentha logifolia (local name Pooneh). No significant correlation was found between seropositivity to fasciolosis and sex, age, history of consumption of green leafy aquatic plants whereas correlation between seropositivity and abdominal pain was significant (P< 0.05). CONCLUSION: Results of this study showed that the seroprevalence rate of human fasciolosis in Yasuj district is relatively high and this area can be considered as a new emerging focus of the disease in Iran.

Expression and Purification of P43 Toxoplasma gondii Surface Antigen.

BACKGROUND: Toxoplasma gondii is an obligate intracellular protozoan parasite, capable of infecting all species of mammals including man. Congenital toxoplasmosis is more important during pregnancy for the first time. In this study we expressed and purified P43 Toxoplasma gondii tachyzoite and bradyzoite specific surface antigen. METHODS: The recombinant pGEMEX-1 contained Toxoplasma P43 coding sequence was transformed into E. coli and mass cultured in LB medium contained 100 µg/ml ampicillin at 37°C over night. The T7 promoter was induced by 1mM isopropyl-1-thio-ß-D-galactopyranoside (IPTG. Recombinant protein was purified by affinity chromatography and confirmed by gel diffusion dot blot and western blot,-using specific anti Toxoplasma antibodies. RESULTS: Recombinant plasmid was induced by IPTG and analyzed by SDS-PAGE. Recombinant protein was confirmed by Western-blot and dot blot using anti human Toxoplasma antibody. CONCLUSION: Recombinant Toxoplasma P43 was produced successfully.

Comparison of Agar Dilution, Broth Dilution, Cylinder Plate and Disk Diffusion Methods for Evaluation of Anti-leishmanial Drugs on Leishmania promastigotes.

BACKGROUND: Leishmaniases are a group of diseases caused by Leishmania parasites. Growing of drug unresponsiveness in leishmaniasis patients necessitates the development of new drugs and accordingly a suitable assay is needed for evaluation of any modalities. The aim of this study was to compare four drug assays methods, agar dilution, broth dilution, cylinder plate and disk diffusion, for evaluation of anti-leishmanial drugs on Leishmania promastigotes, using glucantime as a currently available drug for treatment of leishmaniasis. METHODS: For broth dilution method, different concentration of glucantime was added to the parasite culture (promastigotes of Leishmania), while in cylinder plate method wells were punched in agar gel and filled with different concentration of drug and zone of inhibition was measured in each well. In disk diffusion method, the parasites were cultivated on the surface of agar; filter paper disks were enriched with various concentration of glucantime and were placed on the surface of agar. In agar dilution method, various concentrations of drug were incorporated onto blood agar and the parasites were cultivated on the surface of the agar. RESULTS: A direct correlation was found between the drug concentration and size of inhibitory zones in cylinder plate and disk diffusion methods. These two drug assays methods provided much better performance in comparison with broth and agar dilution methods. CONCLUSION: Cylinder plate and disk diffusion methods seem to be acceptable methods for susceptibility testing of anti-leishmanial compounds on Leishmania promastigotes.

Genotyping of Giardia lamblia isolates from human in southern Iran.

Giardia lamblia cysts isolated from human faeces in South of Iran were analyzed with PCR-restriction fragment length polymorphism (RFLP) assay, based on the detection of glutamate dehydrogenase (gdh) genes. Among 205 faecal samples from microscopically diagnosed giardiasis patients, the gdh gene was amplified from 172 cases with a semi nested PCR assay and typed by RFLP analysis. Of the 172 positive samples, 128 (74.41%) were typed as assemblage AII, 30 (17.44%) assemblage BIII, 6 (3.49%) assemblage BIV and in 8 (4.66%) isolates, mixed assemblages AII and BIV were detected. Clinical features were available for 52 successfully typed cases and the possible correlation of Giardia assemblages and clinical symptoms was evaluated. Both assemblages caused similar illness, but assemblage AII was significantly more frequently associated with abdominal pain, nausea and vomiting. Since these isolates, A and B, are of human origin, anthroponotic transmission of Giardia can be suggested for the route of infection in this region.

Glucantime efficacy in the treatment of zoonotic cutaneous leishmaniasis.

Pentavalent antimony (SbV) compounds are still considered the first line of treatment for all forms of leishmaniasis. There have been reports of drug resistance and unresponsiveness to treatment with these drugs. We investigated the clinical response to treatment of cutaneous leishmaniasis with glucantime, the drug of choice for all forms of leishmaniasis in Iran. All individuals suspected of cutaneous leishmaniasis from October 2007 to March 2008 were included in the study if met specific criteria. After laboratory diagnosis and parasite identification by PCR, 43 patients agreed to participate and complete the protocol for treatment. Meglumine antimoniate (glucantime) was given at a dose of 20 mg/kg/day for 20 days (two 10-day periods) according to a World Health Organization (WHO) recommended protocol. Response to treatment was evaluated 6 weeks after initiation of treatment. Fifteen patients (34.9%) were clinically unresponsive to glucantime treatment while the remaining 28 patients (65.1%) responded to treatment. There were no statistically significant differences by occupation, gender, chronicity of the disease before starting treatment, number of lesions, or age between the glucantime sensitive and resistant patients. Our study showed a significant level of unresponsiveness to glucantime among patients with cutaneous leishmaniasis caused by Leishmania major in Iran. These findings highlight the need for new treatment regimens.

Characterization of the metacaspase 1 gene in Plasmodium vivax field isolates from southern Iran and Italian imported cases.

Plasmodium vivax is still the more prevalent human Plasmodium outside Africa and despite this fact, there is still a deep lack of knowledge on its biology. Metacaspases are cysteine proteases related to metazoan caspases, involved in programmed cell death. Here, we have characterized the P. vivax metacaspase 1 gene in a total of 63 vivax isolates, 32 isolates collected in southern Iran and 31 Italian imported isolates originating from 12 different endemic countries. We have firstly identified DNA size polymorphism in P. vivax metacaspase 1 gene. A total of four different allelic sizes were found, resulting from the insertion of 1 to 4 tandem repeat units located within the intronic region of the P. vivax metacaspase 1. Similarly, we also have identified four distinct allelic types by using vivax merozoite surface protein-1 size polymorphism analysis.

Performance of antigen B isolated from different hosts and cyst locations in diagnosis of cystic echinococcosis.

BACKGROUND: The aim of this study was to assess the performance of Antigen B (AgB) isolated from different Echinococcusgranulosus intermediate hosts and from different cyst locations for immunodiagnosis of human cystic echinococcosis (CE). METHODS: Hydatid cyst fluids were collected from lung and liver cysts of sheep, liver cysts of goats, lung cysts of camels and cattle, and liver cysts of human. AgB was purified from each of these hydatid cysts fluids. Serum samples obtained from 47 pathologically confirmed cases of CE along with 30 sera samples from non-CE patients and 40 sera from healthy individuals were tested by ELISA using AgB prepared from different hosts or cyst locations. RESULTS: The highest sensitivity (97.8%) for diagnosis of CE was seen with AgB prepared from human liver cysts. This maximal sensitivity was followed by AgB isolated from those of sheep liver and lung cysts. The least sensitivity was found with AgB prepared from bovine lung cysts. The highest specificities (97.1%) were observed with AgB isolated from human liver cysts followed by those of sheep and goat liver cysts while the lowest specificity was seen with AgB isolated from bovine lung cysts. In view of the specificities and sensitivities of the different AgB, the best validity was found for AgB prepared from human liver cysts while the least validity was found with AgB prepared from bovine lung cysts. CONCLUSION: For any AgB-based tests, obtaining of the antigen from one of these sources will significantly increase the diagnostic sensitivity and specificity of the assay.

The changing profile of cutaneous leishmaniasis in a focus of the disease in Jahrom district, southern Iran.

Human cutaneous leishmaniasis (CL) is endemic in several parts of Iran, and there is an urban focus of the disease in the district of Jahrom, which forms part of the southern province of Fars. To explore the current profile of the disease in Jahrom district, samples were taken from the skin lesions of 40 cases of CL patients in the district, so that the causative parasites could be identified, to species, in a nested PCR. Although Leishmania tropica has been identified, in the past, as the cause of most of the urban CL in Fars province, the predominant species represented in the recent samples from Jahrom district was L. major (87.5%), while L. tropica was relatively rare (12.5%). More than one in every three (35%) of the cases examined was a child aged <10 years. The most common location of the skin lesions was on the hands. Although most of the cases had one skin lesion each, two cases each had >25 such lesions. The change in the predominant parasite causing CL in Jahrom district, from the L. tropica usually associated with the urban disease in Iran to the L. major more usually associated with CL in rural settings, may well necessitate changes in the local strategies for the prevention and control of CL.

Seroepidemiological study of visceral leishmaniasis in Booyerahmad district, south-west Islamic Republic of Iran.

Visceral leishmaniasis (VL) is endemic in parts of Islamic Republic of Iran. A cross-sectional seroprevalence study was carried out in children in Booyerahmad district in the south-west of the country. Serum samples were taken from 1628 children up to 10 years old from different areas in Booyerahmad in 2005-06. AntiLeishmania antibody was detected in 50 out of 1628 children (3.1%) by direct agglutination test (antibody titre > or = 1:3200). There was no significant difference in seropositivity between the sexes (2.8% males and 3.3% females). The highest rate of infection (5.2%) was in the age group 10 years. Further studies are needed to explore the reservoirs and vectors of the disease in this region.

Comparison of three methods for diagnosis of cutaneous leishmaniasis.

BACKGROUND: Leishmaniasis is one of the infectious parasitic diseases of highest incidence in the world. Cutaneous Leishmaniasis (CL) has long been reported in Shiraz, Southern Iran. There is a need to find a sensitive and specific method for treatment and control of the disease. METHODS: We have compared the sensitivity of the conventional methods microscopy and cultivation of lesion scrapes against PCR amplification of parasite kinetoplast DNA from these samples. The samples (n=219) were obtained from the patients clinically suspected of CL. The smears were stained with Giemsa for microscopy and cultured in Novy-Nicolle-McNeal (NNN) blood agar for promastigote growth. For PCR, the dry smears were scraped off the slides and DNA was extracted. RESULTS: The positive rates from 219 specimens were 76.71%, 50.68%, and 93.61% for microscopy, cultivation, and PCR, respectively. The highest correlation was found between PCR and microscopy method (P=0.014). In PCR assay, 95.61%, 3.9%, and 0.49% of the samples were identified as Leishmania major, L. tropica, and dermatropic L. infantum, respectively. CONCLUSION: The PCR method appears to be the most sensitive for the diagnosis of CL and is valuable for identifying the other species of Leishmania with confusing dermatropic signs.