Cutaneous Leishmaniasis Species Research Articles

A review of the effect of medicinal plant extracts on Leishmania major in vitro and in vivo conditions in Iran

Background: Leishmaniasis is one of the most common parasitic diseases in tropical and subtropical regions of the world and is considered a threat to public health. Iran is also one of the most endemic areas of cutaneous leishmaniasis in the world. The causative species of cutaneous leishmaniasis is a protozoan from the Kinetoplastida order, which in Iran is Leishmania major (rural type) and Leishmania tropica (urban type). More than 70% of leishmaniasis in Iran is Leishmania major. The reservoir of the disease is humans in the urban type and field rodents in the rural type, and the vector is the female mosquito of the genus Phlebotomus. Methods: This study is organized as a review, in which, by searching the keywords cutaneous leishmaniasis, Leishmania major, plants effective against leishmaniasis in Iran, scientific-research articles, Google Scholar search engine information, Pubmed and Science Direct databases, the available books in this field were analyzed. Results: 5-valent antimoan compounds are used for the treatment of cutaneous leishmaniasis. Of course, not all patients need treatment because, in a large number of people, the lesion heals by itself, and due to the side effects of antimoan compounds, it is better to use fewer of these drugs. Therefore, the desire of patients and therapists to use herbal compounds has increased. Conclusion: Although cutaneous leishmaniasis is not usually associated with high Although cutaneous leishmaniasis is not usually associated with high mortality, the rate of infection is very high and causes malformed skin lesions that remain for more than a year in some cases, and even with standard treatment, scars remain. It remains forever and causes emotional pain for the patient. Therefore, domestic researchers have provided research on herbal treatments against Leishmaniasis, considering the history of traditional treatments in Iran and the scattered vegetation in the country.

Molecular Identification of Cutaneous Leishmaniasis Species in the Northcentral Iranian Province of Alborz: Is There a New Focus on Cutaneous Leishmaniasis in the Province?

Cutaneous leishmaniasis (CL) is an endemic infection in the Middle East, including Iran that is also spreading to new foci. We aimed to determine the leishmaniasis species causing CL in Alborz province. Overall, out of 55-suspected CL patients referred to health centers in Alborz Province, north central Iran in 2019, 40 patients had positive smear for CL based on optical microscopy. The internal transcribed spacer 1 (ITS1) of nuclear ribosomal DNA (rDNA) was amplified by PCR. Leishmania species were identified by PCR-restriction fragment length polymorphism (PCR-RFLP) using BshF I (Hae III) enzyme. Out of the 40 positive patients with CL, 34 cases (85%) had been caused by Leishmania (L) major and six (15%) by L. tropica. Fifteen patients had no history of traveling to the disease endemic areas, of which nine were Iranians. Skin lesions and scars caused by CL were mostly observed on the hands and face. Moreover, more than two skin lesions were observed in 22 cases (55%), all of which were infected with L. major. A single skin ulcer was seen in 18 (45%) of the CL patients. Climate change, reduced rainfall, and demographic changes such as migration into Alborz Province and the increasing marginalization of the population and their entry to settle in new areas might have caused natural transmission of both L. tropica and L. major in this province.

Definition of the main vector of cutaneous leishmaniasis: Ecology and mapping in endemic area of Northeast Brazil

Cutaneous leishmaniasis is endemic in Pernambuco. Aiming to determine the vector species of cutaneous leishmaniasis in an endemic area of the Northeast region of Brazil, this study aimed to use the spatial mapping of human cases of CL and correlate with ecological studies of the vectors in the municipality of Timbaúba, Pernambuco, Brazil. Individuals infected with CL were recruited through active search in their homes and clinically and serologically diagnosed during the period from 2018 to 2019. Sandflies were captured with CDC-type light traps in peridomiciliary environments and these were identified at the species level. Females were separated for DNA extraction and subsequent analysis by PCR. The points of collection of phlebotomes and the residences of individuals with lesions were marked with GPS. During the study period, 60 cases of CL were diagnosed. A higher concentration of CL cases was observed in proximity to Atlantic forest remnants confirmed by heat map. A total of 3744 sandflies was captured and five distinct species were identified, with the predominance of Nyssomyia whitmani. From the females separated for the identification of Leishmania braziliensis DNA, a rate of 0.68% of infected sandflies was obtained. It was concluded that cutaneous leishmaniasis continues to be a rural feature of the area. And from this study, it is concluded that Ny. whitmani is the carrier species of CL in the municipality of Timbaúba, Pernambuco. This is due to abundance in catching, specialization of species and PCR positivity for Leishmania braziliensis.

Molecular Identification of Leishmania Species in an Outbreak of Re-Emerged Cutaneous Leishmaniasis in Southwestern Iran During 2015 - 2016

Background: Dezful and its suburbs, as the second city in Khuzestan Province, southwest of Iran, has been an endemic area of cutaneous leishmaniasis (CL) with a low incidence rate since the last decades. However, the disease incidence has rapidly increased, and now is considered as a re-emerging parasitic disease in the area. Objectives: The aim of this study was to identify the most prevalent CL species in Dezful Region. Methods: A total of 196 microscopically confirmed slides from CL patients referred to Dezful Health Center were randomly collected in the period of 2015 - 2016. After DNA extraction from microscopically positive slides, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was carried out on 61 eligible specimens using ribosomal internal transcribed spacer 1 (ITS1) gene. The HaeIII restriction enzyme was used for the identification of species. Results: Samples were randomly collected from 196 acute CL cases, including 110 (56.2%) males and 86 (43.8%) females. Most infections were seen in the age range of 2 - 7 years (65/196, 33.1%). Totally, 60.1% of the cases had only one lesion, and half of the lesions appeared on hands. Furthermore, 162 (82.7%) cases were referred to Dezful Health Center in the cold seasons (autumn and winter). Results of PCR-RFLP on 61 eligible isolates showed that 60 (98.4%) isolates were Leishmania major, and only 1 (1.6%) isolate was Leishmania tropica. Conclusions: Our findings indicated that L. major is the main agent of re-emerged CL in Dezful and its suburbs, and the disease is a zoonosis.

Molecular Identification of Leishmania Species in Patient with Cutaneous Leishmaniasis in an Endemic Area of Fars Province, Southern Iran

The Cutaneous Leishmaniasis (CL) contains a range of factors, which provide different patterns of transmission that make it difficult to control. The aim of this study was to identify the different species of Leishmania in the Jahrom district of Fars province, southern Iran. This descriptive study was done on CL suspected cases who were referential to the Reference of Jahrom Health Center laboratory in 2017 from all different zones mentioned. Demographic variables (age, sex, location, number and place of wounds) was recorded by a questionnaire. The blood smears were checked for the existence of amastigotes by light microscopy. After microscopic examination smears were extract DNA and used in the Polymerase Chain Reaction (PCR) method. Totally 165 slide samples checked were found to be positive for amastigotes via microscopy and positive for leishmanial DNA in the PCR. two (1.2%) of the slide samples gave the 750-bp indicative of Leishmania tropica and 163 samples (98.8%) gave the 560-bp indicative of Leishmania major. Although the actually that both species of CL are common in Iran, but our results showed that in Jahrom district the main specie was L. major. Thus, this fact displays the importance of reservoir rodents in programs of CL control in this County.

Molecular Study of Cutaneous Leishmaniasis Species among Soldiers with Dermal Ulcers in Zahedan, Iran

Molecular Study of Cutaneous Leishmaniasis Species among Soldiers with Dermal Ulcers in Zahedan, Iran

Molecular identification of cutaneous leishmaniasis species using kinetoplast DNA (kDNA) gene in southeast of Iran

Cutaneous leishmaniasis (CL) is one of the most common parasitical diseases in many regions of Iran and plays a significant role in the inhibition of socioeconomic growth of this deprived province. The present study aimed to molecular identification of cutaneous leishmaniasis species in patients from Chabahar and Konark district, in Sistan and Baluchestan province, Iran. This descriptive cross-sectional study was carried out on 250 patients suspected of cutaneous leishmaniasis (CL). Samples of the skin lesions and ulcers suspected to CL were collected from patients and diagnosed by parasitological test and molecular methods, respectively. The microscopic test detected 80 out of 250 (32%) suspected patients to CL, while 80 out of 80 (100%) were positive for Leishmania kDNA. Two species L. major and L. tropica were responsible for 9 and 69 cases, respectively. Our results were consistent with the previous reports in Iran, revealing L. tropica as the most causative agent of CL Further investigation are demanded to identify the hosts, reservoirs, disease species and vectors in this part of the world.

Topical photodynamic therapy with chloroaluminum phthalocyanine liposomes is as effective as systemic pentavalent antimony in the treatment of experimental cutaneous leishmaniasis.

In the Americas, one of the main causative species of cutaneous leishmaniasis is Leishmania (Leishmania) amazonensis. The systemic antimonials remain the most largely used option for disease control. However, this drug has significant toxicity. The development of new alternative therapies, including the identification of effective drugs for topical treatment of cutaneous leishmaniasis, is of utmost interest. In this sense, photodynamic therapy emerges as a new strategy. The aim of this study was to develop the chloroaluminum phthalocyanine-loaded liposome, characterize it, and evaluate its stability and efficacy in the topical treatment of cutaneous leishmaniasis caused by L. (L.) amazonensis. Liposomes composed of egg phosphatidylcholine were prepared by Bangham's method. Storage stability of phthalocyanine-loaded liposomes was evaluated at 30 and 60 days after preparation. For the in vivo evaluation, the animals were infected with L. (L.) amazonensis and divided into groups: chloroaluminium phthalocyanine-loaded liposome, blank liposome, meglumine antimoniate (200 mgSb+5/Kg/day), and control. The lesion size was determined weekly after the beginning of the treatment. Upon completion, parasites were recovered from the skin lesion and spleen and evaluated by limiting dilution assay. Chloroaluminum phthalocyanine-loaded liposomes were stable and showed adequate characteristics for topical administration. The topical chloroaluminum phthalocyanine-loaded liposome was as effective as systemic pentavalent antimony in reducing the parasitic load in the lesion and spleen in infected animals. The present study showed that photodynamic therapy with chloroaluminum phthalocyanine-loaded liposomes is a promising strategy for the treatment of American cutaneous leishmaniasis caused by L. (L.) amazonensis.

Molecular Diagnosis and Phylogenetic Analysis of 5.8s rDNA Gene of Cutaneous Leishmaniasis Species in Holy Karbala/Iraq

Molecular Diagnosis and Phylogenetic Analysis of 5.8s rDNA Gene of Cutaneous Leishmaniasis Species in Holy Karbala/Iraq

Cutaneous Leishmaniasis in Erbil Governorate: Clinical manifestations and disease course.

Background: Cutaneous leishmaniasis (CL) is an infectious disease of zoonotic characteristic; its etiological agent is a protozoan of Leishmania species. This infectious agent is transmitted to humans through a secondary vector, which is infected sand-fly. According to WHO report, Iraq is a country where cutaneous leishmaniasis is an emerging disease, especially, its rural areas are labeled to be the source of the infection and endemicity. Course of the disease and treatment is highly defined by the subtype of cutaneous leishmaniasis and its species. Methods: Data were collected from cutaneous leishmaniasis registry database of Erbil Dermatology Teaching Center in city of Erbil from August 2016 to August 2017. The collected data were descriptively analyzed for the clinical manifestations and the course of disease in the outbreak of this disease in Erbil governorate. Results: A total of 124 patients were enrolled in this study. Among them, 93 (75%) were male and 31 (25%) were female. Their age ranged from 2 years to 73 years, with the mean age of 30.7±11.3. The majority of the patients were members of army forces. The total number of the lesions was 325. The number of lesions being ulcerated at the time of presentation was 179 (55%) and non-ulcerated lesions 146 (45%). Time needed for improvement ranged from 1.5 to 4.8 months. Recovery time of cutaneous leishmaniasis lesions from time of onset to the remission ranged from 1.3 to 14.3 months. Conclusion: The complete recovery time of the CL lesions was longer than what has been mentioned previously in Iraq. Ulceration of the lesions depends on the diameter of the lesions, as increased diameter increases the possibility of the ulceration. The possibility of lesion ulceration increases with an increase in the diameter of the lesions. Considering the observations in different clinical patterns and course of this disease, further studies should be conducted to identify CL species.

Geographical distribution and molecular characterization for cutaneous leishmaniasis species by sequencing and phylogenetic analyses of kDNA and ITS1 loci markers in south-eastern Iran

This study aimed to explore geographic distribution and molecular characterization of cutaneous leishmaniasis (CL) species by amplifying two popular markers in kinetoplast DNA and internal transcribed spacer 1 loci by nested-PCR, and characterized by sequencing and phylogenetic analyses. Findings demonstrated that two species co-existed in the province: L. tropica (88.5%) and L. major (11.5%). All gender and age groups were equally infected, although males, 21–30 years old, exhibited a significantly higher infection. Sequencing and phylogenetic analyses of 34 randomly selected samples showed that L. tropica isolates exhibited some degree of heterogeneity. Both anthroponotic CL and zoonotic CL are present in south-eastern Iran with predominance of L. tropica species. Some level of heterogeneity was observed in L. tropica isolates which possibly reflects different colonies in the area. Implementation of diagnostic tools directly from clinical samples could be an important strategic approach for exploration of spatial distribution, molecular characterization and phylogenetic analyses.

Leishmania major cutaneous leishmaniasis outbreak in the Jordanian side of the Northern Jordan Valley

Cutaneous leishmaniasis (CL) is endemic in many foci of Jordan and the Jordanian Mid Jordan Valley (JMJV) is the most affected and the incidence is quite high. The situation in the northern part of the Jordanian side of the Jordan Valley (NJJV) was different; before 2008, CL has rarely been reported from this area. From April 2008 to May 2009, passive detection followed by active detection was used to trace cases of CL from the NJJV. DNA was extracted from seven clinical isolates of Leishmania promastigotes and lesion scrapings spotted on filter papers obtained from 51 suspected CL patients living in the NJJV. The identity of the causative species of CL in the NJJV was investigated using ITS1-PCR followed by RFLP. In 2008/2009, 183 cases were clinically diagnosed of having CL in the NJJV. The parasites in five of the isolates and in 48 PCR-positive scrapings were classified as Leishmania major. In two isolates and in one PCR-positive scraping Leishmania tropica was identified. Investigations on the origin of CL cases revealed that the L. tropica cases were residents of two towns outside the NJJV. Herein, we report the clinical features, parasitological diagnosis, etiology, and the geographical distribution of CL cases from NJJV with the aim of documenting, for the first time, an outbreak in this area.

Viscerotropic leishmaniasis: a systematic review of the case reports to highlight spectrum of the infection in endemic countries

AbstractVisceral leishmaniasis is an important neglected parasitic disease that is generally caused byLeishmania infantum,Leishmania donovaniandLeishmania chagasi. However, several causative species of cutaneous leishmaniasis (CL) causes an interstitial form of leishmaniasis which known viscerotropic leishmaniasis. The aim of this paper is a systematic review of the cases of viscerotropic leishmaniasis to present the main causative agents, clinical manifestations, treatment and outcomes of the cases. An electronic search (any date to August 2017) without language restrictions was performed using Medline, PubMed, Scopus and Google Scholar. The searches identified 19 articles with total 30 case reports. Of them, old worldLeishmaniaspecies was reported from 23 (76.7%) cases, including 20 cases ofL. tropicaand three cases ofL. major; whereas new worldLeishmaniaspecies were reported in seven (23.4%) cases. The infection was more prevalent in male (24/30, 80%) than female (5/30, 16.7%) patients. Co-morbidity/co-infection was observed in 13 out of 30 cases (43.4%), which the most of them was HIV/AIDS (10 out of 13 cases, 76.9%). The results suggested that viscerotropic leishmaniasis should be more attended in the endemic countries of CL and in immunocompromised patients in order to exact discrimination from other endemic infectious diseases.

Curvas de fusión de regiones genómicas específicas: una herramienta prometedora para el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia.

Introducción. La leishmaniasis cutánea es una enfermedad causada por parásitos del género Leishmania que tiene gran incidencia en Colombia. El diagnóstico y la identificación de la especie infecciosa son factores críticos en el momento de escoger e iniciar el tratamiento. Actualmente, los métodos de diagnóstico y tipificación requieren procedimientos complejos, por lo que es necesario validar nuevos marcadores moleculares y métodos que simplifiquen el proceso.Objetivo. Desarrollar una herramienta basada en la reacción en cadena de la polimerasa (PCR) con curvas de fusión (High Resolution Melting; PCR-HRM) para el diagnóstico y tipificación de las tres especies de Leishmania de importancia epidemiológica en casos de leishmaniasis cutánea en Colombia.Materiales y métodos. Los genomas de Leishmania panamensis, L. braziliensis y L. guyanensis se compararon mediante métodos bioinformáticos. Las regiones específicas de especie identificadas se validaron mediante PCR. Para los marcadores seleccionados se diseñó una PCR-HRM y se estimaron algunos parámetros de validez y seguridad usando aislamientos de pacientes colombianos caracterizados previamente mediante PCR y análisis de polimorfismos en la longitud de los fragmentos de restricción (Restriction Fragment Length Polymorphism - RFLP; PCR-RFLP) del gen hsp70.Resultados. El análisis genómico comparativo mostró 24 regiones específicas de especie. Sin embargo, la validación mediante PCR solo identificó un marcador específico para cada especie de Leishmania. Los otros marcadores mostraron amplificación cruzada. El límite de detección para los tres marcadores seleccionados fue de un parásito, mientras que la sensibilidad, la especificidad, el valor predictivo positivo y el negativo fueron de 91,4, 100, 100 y 75 %, respectivamente.Conclusiones. Las tres regiones seleccionadas pueden emplearse como marcadores moleculares en el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia.

Identification of Leishmania Species for Cutaneous leishmaniasis in Gonabad, Bardaskan and Kashmar, Central Khorasan, 2015

Background: About 90% of cutaneous leishmaniasis (CL) cases are reported from 7 countries including Iran. In this study, the cutaneous leishmaniasis species causing CL in Gonabad, Bardaskan, Kashmar cities in central Khorasan (Iran) were identified by kDNA-PCR. Methods: During the study, 93 suspected patients with CL, who were referred to the dermatology research center in these cities, were evaluated based on age, clinical forms, and place of residence. Direct microscopy was employed for parasitological diagnosis and PCR from skin ulcers performed using specific kDNA primers. Data were analyzed using the SPSS software. Results: Among 93 individuals with skin ulcers suspected to CL, the results of 81 direct smears were positive. PCR bands were observed in 84 examined samples, of which 68 (81%) samples were identified for Leishmania tropica and 16 (19%) samples for L. major. 12 patients were from Bardaskan with 10 cases of L. tropica, 23 patients from Gonabad with 20 cases of L. tropica, and 49 patients from Kashmar with 38 cases of L. tropica. Most of the lesions were located on hands (37%), the most clinical feature was papule (75%), and most of patients were 21 - 30 years old. Conclusions: Previous epidemiologic studies have indicated that anthroponotic cutaneous leishmaniasis is the only dominant CL in the center and south of Khorasan. However, this study introduced new foci of zoonotic cutaneous leishmaniasis in these areas. Kashmar city was introduced as a new focus of zoonotic cutaneous leishmaniasis.

Cutaneous Leishmaniasis in Khyber Pakhtunkhwa Province of Pakistan: Clinical Diversity and Species-Level Diagnosis.

This study primarily aimed to identify the causative species of cutaneous leishmaniasis (CL) in the Khyber Pakhtunkhwa Province of Pakistan and to distinguish any species-specific variation in clinical manifestation of CL. Diagnostic performance of different techniques for identifying CL was assessed. Isolates of Leishmania spp. were detected by in vitro culture, polymerase chain reaction (PCR) on DNA extracted from dried filter papers and microscopic examination of direct lesion smears from patients visiting three major primary care hospitals in Peshawar. A total of 125 CL patients were evaluated. Many acquired the disease from Peshawar and the neighboring tribal area of Khyber Agency. Military personnel acquired CL while deployed in north and south Waziristan. Leishmania tropica was identified as the predominant infecting organism in this study (89.2%) followed by Leishmania major (6.8%) and, unexpectedly, Leishmania infantum (4.1%). These were the first reported cases of CL caused by L. infantum in Pakistan. PCR diagnosis targeting kinetoplast DNA was the most sensitive diagnostic method, identifying 86.5% of all samples found positive by any other method. Other methods were as follows: ribosomal DNA PCR (78.4%), internal transcribed spacer 2 region PCR (70.3%), culture (67.1%), and microscopy (60.5%). Clinical examination reported 14 atypical forms of CL. Atypical lesions were not significantly associated with the infecting Leishmania species, nor with “dry” or “wet” appearance of lesions. Findings from this study provide a platform for species typing of CL patients in Pakistan, utilizing a combination of in vitro culture and molecular diagnostics. Moreover, the clinical diversity described herein can benefit clinicians in devising differential diagnosis of the disease.

Geographic Distribution of Leishmania Species in Ecuador Based on the Cytochrome B Gene Sequence Analysis.

A countrywide epidemiological study was performed to elucidate the current geographic distribution of causative species of cutaneous leishmaniasis (CL) in Ecuador by using FTA card-spotted samples and smear slides as DNA sources. Putative Leishmania in 165 samples collected from patients with CL in 16 provinces of Ecuador were examined at the species level based on the cytochrome b gene sequence analysis. Of these, 125 samples were successfully identified as Leishmania (Viannia) guyanensis, L. (V.) braziliensis, L. (V.) naiffi, L. (V.) lainsoni, and L. (Leishmania) mexicana. Two dominant species, L. (V.) guyanensis and L. (V.) braziliensis, were widely distributed in Pacific coast subtropical and Amazonian tropical areas, respectively. Recently reported L. (V.) naiffi and L. (V.) lainsoni were identified in Amazonian areas, and L. (L.) mexicana was identified in an Andean highland area. Importantly, the present study demonstrated that cases of L. (V.) braziliensis infection are increasing in Pacific coast areas.

Real-time PCR using FRET technology for Old World cutaneous leishmaniasis species differentiation.

BackgroundRecently, there has been a re-emergence of cutaneous leishmaniasis in endemic countries and an increase in imported cases in non-endemic countries by travelers, workers, expatriates, immigrants, and military force personnel. Old World cutaneous leishmaniasis is caused primarily by Leishmania major, L. tropica and L. aethiopica. Despite their low sensitivity, diagnosis traditionally includes microscopic and histopathological examinations, and in vitro cultivation. Several conventional PCR techniques have been developed for species identification, which are time-consuming and labour-intensive. Real-time PCR using SYBR green dye, although provides rapid detection, may generate false positive signals. Therefore, a rapid and easy method such as a FRET-based real-time PCR would improve not only the turn-around time of diagnosing Old World cutaneous Leishmania species but will also increase its specificity and sensitivity.MethodsA FRET-based real-time PCR assay which amplifies the cathepsin L-like cysteine protease B gene encoding a major Leishmania antigen was developed to differentiate L. major, L. tropica, and L. aethiopica in one single step using one set of primers and probes. Assay performance was tested on cutaneous and visceral strains of Leishmania parasite cultures and isolates of other protozoan parasites as well as human biopsy specimen.ResultsThe assay readily differentiates between the three Old World cutaneous leishmaniasis species based on their melting curve characteristics. A single Tm at 55.2 ± 0.5 °C for L. aethiopica strains was distinguished from a single Tm at 57.4 ± 0.2 °C for L. major strains. A double curve with melting peaks at 66.6 ± 0.1 °C and 48.1 ± 0.5 °C or 55.8 ± 0.6 °C was observed for all L. tropica strains. The assay was further tested on biopsy specimens, which showed 100 % agreement with results obtained from isoenzyme electrophoresis and Sanger sequencing.ConclusionCurrently, there are no published data on real-time PCR using FRET technology to differentiate between Old World cutaneous Leishmania species. In summary, our assay based on specific hybridization addresses the limitations of previous PCR technology and provides a single step, reliable method of species identification and rapid diagnostic applications.

Isolation and identification of cutaneous leishmaniasis species by PCR-RFLP in Ilam province, the west of Iran.

Cutaneous leishmaniasis (CL) is one of the most common parasitic diseases and public health problems in Iran. CL is endemic in most parts of Ilam province, in the west of Iran. The distance from the center of country, the great number of divers rural areas, and lack of specialists and laboratory facilities have been the major causes of Leishmania species remaining unknown in this region. Polymerase chain reaction followed by restriction fragment length polymorphism was performed to identify the Leishmania species in 61 patients with cutaneous lesions. Eventually L. major was confirmed as the cause of cutaneous leishmaniasis in Ilam province, the west of Iran.

The Jordanian Mid Jordan Valley is a classic focus of Leishmania major as revealed by RFLP of 56 isolates and 173 ITS-1-PCR-positive clinical samples

The identity of the causative species of cutaneous leishmaniasis (CL) in the endemic Jordanian Mid Jordan Valley (JMidJV) was investigated using the polymerase chain reaction (PCR) amplifying the ribosomal internal transcribed spacer 1 (ITS-1) followed by the restriction fragment length polymorphism (RFLP). The geographical distribution of CL and the usefulness of ITS1 PCR in diagnosis of suspected CL in the study area were also addressed. Over the period from 2004 to 2009, 56 clinical isolates of Leishmania promastigotes and 185 lesion scrapings spotted on filter papers were obtained from suspected CL patients living in the JMidJV, which is divided into northern and southern districts. The majority (67.1%) of patients occurred in the populated eastern part of the southern district. Of the 185 suspected CL patients, 173 (93.5%) were confirmed positive using PCR. Leishmanial DNA was detected in 27 (90%) of 30 patients having clinically atypical lesions of CL and in 60 (92%) of 65 smear- and culture-negative cases having typical lesions of CL. The parasites in all of the 56 isolates and the 173 PCR-positive scrapings were classified as Leishmania major. In conclusion, PCR is useful in diagnosis of CL especially when smear and culture are negative. It is also recommended as a differential diagnostic tool of atypical lesions when CL is endemic. The identification of L. major as the causative species in such a considerable number of CL cases, representative of all mini foci of CL in the study area, shows that the JMidJV is a classic focus of L. major.