Molecular Diagnosis and Species Identification of Mucosal Leishmaniasis in Iran and Correlation with Cytological Findings

Shirian, Sadegh; Oryan, Ahmad; Hatam, Gholamreza; Daneshbod, Khosrow; Daneshbod, Yahya

doi:10.1159/000337450

Cited by 19 publications

(20 citation statements)

References 35 publications

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“…3 It is a well-known clinical manifestation of infection that is usually manifested after infection with species belonging to the Leishmania (Viannia) subgenus in Central and South America and has occasionally been associated with infections caused by Leishmania species endemic in the Old World. 4,5 Clinically or histologically, ML can be mistaken for benign and malignant lesions. Since cytology is inexpensive and easy to perform, it is the preferred primary approach to identification.…”

Section: Introductionmentioning

confidence: 99%

“…The PCR-based method not only appears to be a useful and more precise diagnostic approach to the identification of suspected cases of ML with negative cytology but is also efficient in determining the species of the parasite. 5 A few ML cases have previously been described in the Old World in infections caused by Leishmania tropica, Leishmania major, and Leishmania infantum, [6][7][8] but no etiological agents of the ML form have been isolated from the patients in Iran. The objective of the present study was to report diagnosis, prognosis, tropism, clinical features and prevalence of three Leishmania species causing the primary and secondary ML in two endemic areas of Iran.…”

Section: Introductionmentioning

confidence: 99%

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ThreeLeishmania/L.species –L. infantum,L. major,L. tropica– as causative agents of mucosal leishmaniasis in Iran

Shirian

Oryan

Hatam

et al. 2013

Pathogens and Global Health

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Cases of human oro-mucosal leishmaniasis are mainly reported in areas where Leishmania (Viannia) braziliensis perpetuates and the damages are mainly located at the cartilaginous nasal septum and frontal portions of the nasal fossa. In Iran, an area free of any L.(V) braziliensis, three Leishmania species are known to perpetuate through distinct (i) blood-feeding sand flies and (ii) rodents or (iii) canidae. Thus while establishing the diagnosis of any human oro-mucosal lesions, three Leishmania species -L. infantum, L. major, and L. tropica -must be considered as potential etiological agents of these damages. With these objectives in mind, features such as localization, extent, severity of oro-mucosal lesions, and duration of symptoms at the time of diagnosis were recorded from 11 patients with respect to the presence or absence of cutaneous lesions in other body parts. The biopsy samples were collected from the oromucosal and cutaneous lesions and were processed for further identification of the Leishmania species. The lesions ranged from mucosal nodules without ulceration, nodules with erosion, and shallow to deep ulcerations. Leishmania major was isolated from six (55%) cases showing lesions or scars. The scars were restricted to upper and lower extremities. For the other five patients who did not display any signs of former or active cutaneous leishmaniasis, L. major, L. tropica, and L. infantum were isolated from their lesions. In conclusion L. major, L. infantum, and L. tropica, regardless of common tropism, can be seen in mucosal tissues. However, L. major was the predominant species detected from the lesions in the nasal, gingival, and hard and soft palates, and L. tropica was isolated from the gingival and lower lip lesions. Leishmania infantum was isolated from two severe cases of deep mucosal damage displayed by the epiglottis, cricoarytenoid muscle, and laryngeal mucosa. One important finding was the association of L. major with active or scarred skin lesions.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

ThreeLeishmania/L.species –L. infantum,L. major,L. tropica– as causative agents of mucosal leishmaniasis in Iran

Shirian

Oryan

Hatam

et al. 2013

Pathogens and Global Health

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“…Mucosal leishmaniasis (ML) is a rare disease in the world, even in areas where it is endemic, such as Iran (6,25). The importance of ML is due to the severity of its clinical lesions, poor response to traditional antimony therapy, and destruction of the nasal architecture with gross facial alterations (17).…”

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confidence: 99%

Mixed Mucosal Leishmaniasis Infection Caused by Leishmania tropica and Leishmania major

et al. 2012

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cMixed infections with different Leishmania species could explain differences in the clinical courses of these infections. On identification of Leishmania parasites from Iranian patients with mucosal leishmaniasis (ML), a patient with both oral and nasal lesions was found to be concomitantly infected with Leishmania tropica and L. major. Mixed infection was identified by PCR amplification of Leishmania kinetoplast DNA on scraping of cytological smears and histopathological sections. L. major and L. tropica were isolated from the nasal and oral lesions, respectively. These species were also confirmed by immunohistochemistry. This seems to be the first reported case of concurrent ML infection with two Leishmania species. It indicates that, at least in this patient, previous infection with one of these Leishmania species did not protect against infection with the other. This result has important implications for the development of vaccines against leishmaniases and implies careful attention in the treatment of this infectious disease. CASE REPORTA 34-year-old immunocompetent male patient presented with lesions of the mucous membranes of the nose and mouth. The patient was from Fars Province, southern Iran. He presented with a 7-and 5-month history of intranasal and oral lesions, respectively. No scar or other lesion was found in other parts of the body. On examination, there were multiple tiny erythematous lesions, varying in size from 0.1 to 0.3 cm in diameter. The nasal pyramid was edematous, and bloody crusts were observed on the inferior conchae, septum, and floor of the nasal fossa. The nasal lesions were located in the intranasal portion in the mucous membrane over the turbinates, far from the cutaneous lesions. Clinically, diffuse yellowish white erosions with grayish fibrinous membranes were seen on a reddish edematous background on the involved oral mucosa (Fig. 1).His blood biochemistry and complete blood count were within the reference range. His hemoglobin level was 13.6 g/dl, his total leukocyte count was 6,600/mm 3 , his serum creatinine level was 0.8 mg/dl, and his blood urea nitrogen level was 18 mg/dl. Serological studies for human immunodeficiency virus and hepatitis B and C viruses were negative.Tissue samples from the lesions were fixed in 10% neutral buffered formalin, embedded in paraffin, sectioned at a 5-m thickness, and stained with hematoxylin and eosin.Cytologic smears were prepared by scraping of the oral lesions with a scalpel. In addition, exfoliative cytology from the nasal lesions was performed by washing the nasal cavity as previously noted (7). Multiple smears were made on slides and were both air dried and alcohol fixed and then stained by the Wright method. Review of the cytologic smears and histologic sections was conducted blindly by three pathologists. Microscopic examination showed the amastigote forms of Leishmania ( Fig. 2A and B).The antibodies IS2-2B4 (A11; specific for L. tropica) and XLVI-5B8-B3 (T1; specific for L. major) were kindly provided by the Special Programme f...

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“…Therefore, the presence of a granulomatous reaction, a high percentage of macrophages and multinucleated giant cells are important suggestive clues in the diagnosis of this disease in cases living in endemic areas, and are clinically suspected to be CL but demonstrate negative histological and cytological manifestations. The presence of a granulomatous reaction and multinucleated giant cells has previously been reported in the negative smears of the patients who showed a clinical manifestation of mucosal leishmaniasis [29,30]. …”

Section: Discussionmentioning

confidence: 94%

Correlation of Genetic Heterogeneity with Cytopathological and Epidemiological Findings of <b><i>Leishmania major</i></b> Isolated from Cutaneous Leishmaniasis in Southern Iran

Shirian

Oryan²,

Hatam³

et al. 2016

Acta Cytologica

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Objective: Cutaneous leishmaniasis (CL) has recently grown into a major public health problem in 88 countries of the world, including Iran. It is a polymorphic disease which may show various clinicopathological features. Although the effect of the genetic diversity of the parasite has been demonstrated as one of the factors influencing clinical manifestations in CL, no data exist regarding the genetic variation of Leishmania major and its microscopical features. Study Design: Fine-needle aspiration, touch smears and the histological sections of 100 patients were examined for Leishmania amastigotes, using Giemsa and hematoxylin and eosin. Diverse types of inflammatory cells in the 40 positive and 5 negative smears were differentiated. Kinetoplast DNA (kDNA) was amplified using nested PCR and subsequently sequenced. Sequencing analysis of the amplified kDNA was used to investigate the genetic variations among L. major isolates and to correlate the findings with microscopical features and geographical origins. Results: The quantified amastigote density in the 40 positive touch smears was blindly classified by 3 observers. Grade I, II, III and IV had 7, 13, 9 and 11 cases, respectively. The microscopical features, the mean percentage of neutrophils, lymphocytes and other inflammatory cells, and the leishmanial density of the grades and negative cytopathological samples were contrasting. kDNA amplification of L. major was detected from the cutaneous lesion, and 21 of these amplicons were successfully sequenced. Conclusions: These results indicate that L. major strains causing CL in southern Iran are genetically diverse; furthermore, a correlation between the genetic heterogeneity of the parasite, the microscopical manifestation and the geographical regions of the disease in humans was found.

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Molecular Diagnosis and Species Identification of Mucosal Leishmaniasis in Iran and Correlation with Cytological Findings

Cited by 19 publications

References 35 publications

ThreeLeishmania/L.species –L. infantum,L. major,L. tropica– as causative agents of mucosal leishmaniasis in Iran

ThreeLeishmania/L.species –L. infantum,L. major,L. tropica– as causative agents of mucosal leishmaniasis in Iran

Mixed Mucosal Leishmaniasis Infection Caused by Leishmania tropica and Leishmania major

Correlation of Genetic Heterogeneity with Cytopathological and Epidemiological Findings of <b><i>Leishmania major</i></b> Isolated from Cutaneous Leishmaniasis in Southern Iran

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