RAPD-PCR reveals genetic polymorphism among Leishmania major strains from Tunisian patients

Yazidi, Rihab; Bettaieb, Jihéne; Ghawar, Wissem; Jaouadi, Kaouther; Châabane, Sana; Zaâtour, Amor; Salah, Afif Ben

doi:10.1186/s12879-015-1010-0

Cited by 6 publications

(5 citation statements)

References 27 publications

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“…In Iran, Oryan et al [33] have reported a high genetic polymorphism of L. major and correlations between the geographical origin and the clinical manifestations of the disease. Nevertheless, Yazidi et al [41] reported that Tunisian L. major isolates are highly polymorphic despite inducing a similar clinical picture and belonging to the same geographical origin.…”

Section: Discussionmentioning

confidence: 99%

Clinical Presentation of Cutaneous Leishmaniasis caused by Leishmania major

Remadi

Haouas²,

Chaara³

et al. 2016

Dermatology

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Background/Aims: The diagnosis of cutaneous leishmaniasis (CL) is based on the microscopic detection of amastigote, isolation of the parasite, or the detection of Leishmania DNA. Nevertheless, since these techniques are time consuming and not usually available in many endemic countries, the diagnosis remains clinical. Consequently, such disease may be overlooked because of its similarity to other skin diseases. The aim of this study is to describe the clinical polymorphism of CL caused by Leishmaniamajor.Methods: A cross-sectional survey was carried out on 166 patients. Diagnoses were made by both microscopic examination of stained tissue-scraping smears and PCR. The Leishmania species was identified by restriction enzyme analysis of the ribosomal internal transcribed spacer 1 region. The clinical polymorphism was analyzed only for patients with a positive diagnosis for CL and L. major as the identified species. Results and Conclusion: Of the 166 patients, 75 patients fit the inclusion criteria. Twelve different types of CL caused by L. major were defined. The most common type was the ulcero-crusted form followed by the papulonodular form and the impetigenous form. The ulcerated, mucocutaneous, lupoid, and sporotricoid forms were less common. The eczematiform, erysipeloid, verrucous, psoriasiform, and pseudotumoral types were represented by a single case. Zoonotic CL caused by L. major can simulate many other skin diseases, which may lead to a significant spread of this disease and increases in morbidity and drug resistance. This large polymorphism may be the result of a complex association between the genetics of the parasite and the immune response of the host.

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Section: Discussionmentioning

confidence: 99%

Clinical Presentation of Cutaneous Leishmaniasis caused by Leishmania major

Remadi

Haouas²,

Chaara³

et al. 2016

Dermatology

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“…Oryan et al [61] and Shirian et al [62] assessed the heterogeneity of L. major causing CL based on sequencing of kDNA and showed a high genetic diversity of the parasite and correlations among the geographical origin and the clinical outcomes of the disease. Moreover, conspicuous genetic variability has been exhibited within the Nagt gene amidst L. tropica , L. major , and L. infantum strains [70, 71], and by RAPD-PCR among L. major and L. infantum strains [72–74]. Considerable genetic diversity was detected among L. major strains from different endemic areas and even between some isolates of the same endemic area in Iran using the RAPD technique [73].…”

Section: Discussionmentioning

confidence: 99%

Leishmania cytochrome b gene sequence polymorphisms in southern Iran: relationships with different cutaneous clinical manifestations

et al. 2019

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BackgroundCutaneous leishmaniasis (CL) caused by Leishmania species, is a geographically extensive disease that infects humans and animals. CL is endemic in half of the 31 provinces of Iran, with 29,201 incidence cases reported in Fars province from 2010 to 2015. CL is polymorphic and may result in lesions characterized by different clinical features. Parasite genetic diversity is proposed to be one of the factors affecting the clinical outcome and lesion characteristics in CL patients. However, there is still very limited data regarding the genetic variation of Leishmania spp. based on the sequencing of Cytochrome b (Cyt b) gene.MethodsAll patients originated from endemic regions in Fars province. The amplification of the Cyt b gene from isolates of 100 patients with disparate clinical forms of CL was accomplished using Nested-PCR. Sequence analysis of the amplified Cyt b was used to scrutinize the genetic variations among Leishmania isolates and connect the results with clinical pictures. The clinical demonstrations were basically of two types, typical and atypical lesions. Molecular phylogenetic tree was constructed using the Neighbor-Joining method, with species/strains from this study compared to species/strains from other geographical regions.ResultsLeishmania major was identified as the predominant infecting Leishmania spp. (86% of cases), with the remainder of cases being infected by Leishmania tropica. Clinical examination of patients revealed 12 different clinical CL forms. Among Leishmania samples analyzed, five distinct haplotypes were recognized: three in L. major and two in L. tropica. We found a correlation between clinical outcomes and Cyt b sequence variation of Leishmania spp. involved. Moreover, we observed a higher presence of polymorphisms in L. major compared with L. tropica. This difference may be due to the different eco-epidemiologies of both species, with L. tropica being an anthroponosis compared to L. major, which is a zoonosis.ConclusionsThe sequence analysis of Cyt b gene from 25 L. major and L. tropica strains demonstrated genetic variability of L. major and L. tropica causing CL in southern Iran, and a feasible connection amid the genetic heterogeneity of the parasite, geographical source and clinical appearance of the disease in human was detected.

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“…The ERIC-PCR products described by Leal (2010) ranged in size from 100 to 1700 bp for Leishmania reference samples (FIOCRUZ) and isolates from 11 patients with visceral leishmaniasis. Yazidi et al (2015) used RAPD-PCR to determine the genetic variability of 39 Leishmania major isolates from five endemic foci. Their analysis revealed a high degree of polymorphism (43%) among isolates, and showed that 99% of the observed variation was within, rather than between, each region studied.…”

Section: Discussionmentioning

confidence: 99%

Standardization of a molecular method for epidemiologic identification of Leishmania strains

et al. 2016

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Molecular studies of the evolutionary relationships among Leishmania species suggest the presence of high genetic variation within this genus, which has a direct effect on public health in many countries. The coexistence of species in a particular region can result in different leishmaniasis clinical forms and treatment responses. We aimed to standardize the kinetoplast DNA (kDNA) enterobacterial repetitive intergenic consensus (ERIC) sequence polymerase chain reaction (PCR) method for molecular epidemiological identification of Leishmania strains, and estimate existing inter-strain genomic differences and kDNA signatures using this technique. ERIC-PCR of genomic DNA revealed genetic polymorphisms between species, although some strains shared many DNA fragments. Leishmania guyanensis, L. amazonensis, and L. braziliensis clustered together in a dendrogram with similarities ranging from 42.0 to 61.0%, whereas L. chagasi grouped with these three species with a similarity of 28.0%. After amplification of kDNA, 780-bp bands were extracted from an agarose gel and purified for analysis of its genetic signature. kDNA ERIC-PCR electrophoretic patterns consisted of 100- to 600- bp fragments. Using these profiles, L. braziliensis and L. guyanensis grouped with a similarity of 26.0%, and L. amazonensis and L. chagasi clustered based on a similarity of 100%. The electrophoretic profiles and dendrograms showed that, for epidemiological identification by ERIC-PCR, genomic DNA had greater discriminatory power than kDNA did. More strains need to be analyzed to validate the kDNA ERIC-PCR method. The genomes of these strains should be sequenced for better epidemiological identification of Leishmania species.

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RAPD-PCR reveals genetic polymorphism among Leishmania major strains from Tunisian patients

Cited by 6 publications

References 27 publications

Clinical Presentation of Cutaneous Leishmaniasis caused by <b><i>Leishmania major</i></b>

Clinical Presentation of Cutaneous Leishmaniasis caused by <b><i>Leishmania major</i></b>

Leishmania cytochrome b gene sequence polymorphisms in southern Iran: relationships with different cutaneous clinical manifestations

Standardization of a molecular method for epidemiologic identification of Leishmania strains

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