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

BMC Infectious Diseases

Afif Ben Salah¹  Amor Zaatour¹  Sana Châabane¹  Kaouther Jaouadi¹  Wissem Ghawar¹  Jihene Bettaieb¹  Rihab Yazidi¹ 

[1] Department of Medical Epidemiology, Laboratory of Transmission, Control and Immunobiology of Infections (LR11IPT02), Pasteur Institute of Tunis, 13 Place Pasteur BP-74, Tunis 1002, Belvedere, Tunisia

关键词: Tunisia;    RAPD-PCR;    Genetic Polymorphism;    Leishmania major;   

Others  :  1230430
DOI  :  10.1186/s12879-015-1010-0

 received in 2014-12-04, accepted in 2015-07-06,  发布年份 2015

【 摘 要 】

Background

Zoonotic cutaneous leishmaniasis caused by Leishmania (L.) major is endemoepidemic in the Center and South of Tunisia. The clinical course of the disease varies widely among different patients and geographic regions. Although genetic diversity in L. major parasites has been suggested as a potential factor influencing their pathogenic variability, little information on genetic polymorphism among L. major strains is available in the literature. This work aimed to estimate the genetic variability within different isolates of L. major.

Methods

Our sample comprised 39 isolates (confirmed as L. major by restriction fragment length polymorphism typing) from patients experiencing the same clinical manifestations but living in different regions of Tunisia where L. major is endemic. Random amplified polymorphic DNA (RAPD) PCR marker polymorphism was estimated by calculating Nei and Li’s genetic distances and by an analysis of molecular variance (AMOVA).

Results

Analysis of the genetic diversity among the isolates revealed a high level of polymorphism (43 %) among them. AMOVA indicated that the highest variability (99 %) existed within the study regions.

Conclusions

Our results revealed a heterogeneous genetic profile for L. major with similar clinical manifestations occurring within the different geographical regions. Additional L. major isolates from patients, insect vectors, and reservoir hosts from different endemic foci should be collected for further analysis.

【 授权许可】

   
2015 Yazidi et al.

【 图 表 】

Fig. 1.

Fig. 2.

【 参考文献 】

• [1]Masmoudi A, Ayadi N, Boudaya S, Meziou TJ, Mseddi M, Marrekchi S et al.. [Clinical polymorphism of cutaneous leishmaniasis in centre and south of Tunisia]. Bull Soc Pathol Exot. 2007; 100:36-40.
• [2]Solbach W, Laskay T. The host response to Leishmania infection. AdvImmunol. 2000; 74:275-317.
• [3]Ashford RW. The leishmaniases as emerging and reemerging zoonoses. Int J Parasitol. 2000; 30:1269-1281.
• [4]Eswarappa SM, Estrela S, Brown SP. Within-host dynamics of multi-species infections: facilitation, competition and virulence. PLoS One. 2012; 7:e38730.
• [5]Waki K, Dutta S, Ray D, Kolli BK, Akman L, Kawazu S et al.. Transmembrane molecules for phylogenetic analyses of pathogenic protists: Leishmania-specific informative sites in hydrophilic loops of trans- endoplasmic reticulum N-acetylglucosamine-1-phosphate transferase. Eukaryot Cell. 2007; 6:198-210.
• [6]Baghaei M. Intraspecific variation in Leishmania major isolated from different forms of zoonotic Cutaneous Leishmaniasis. Iran J Med Sci. 2005; 30:51-54.
• [7]Oryan A, Shirian S, Tabandeh MR, Hatam GR, Randau G, Daneshbod Y. Genetic diversity of Leishmania major strains isolated from different clinical forms of cutaneous leishmaniasis in southern Iran based on minicircle kDNA. Infect Genet Evol. 2013; 19:226-231.
• [8]Cupolillo E, Momen H, Grimaldi G. Genetic diversity in natural populations of New World Leishmania. Mem Inst Oswaldo Cruz. 1998; 93:663-668.
• [9]Haouas N, Gorcii M, Chargui N, Aoun K, Bouratbine A, Messaadi Akrout F et al.. Leishmaniasis in central and southern Tunisia: current geographical distribution of zymodemes. Parasite. 2007; 14:239-246.
• [10]Ikram G, Dellagi K, Ismail RB. Random amplified polymorphic DNA technique for identification and differentiation of old world Leishmania species. Am J Trop Med Hyg. 2002; 66:152-156.
• [11]Hajjaran H, Mohebali M, Razavi MR, Rezaei S, Kazemi B, Edrissian GH et al.. Identification of Leishmania species isolated from human cutaneous leishmaniasis, using random amplified polymorphic DNA, (RAPD-PCR). Iranian Journal of Public Health. 2004; 33:8-15.
• [12]Armstrong JS, Gibbs AJ, Peakall R, Weiler G. The RAPDistance package, ver. 1.03. Research School of Biological Sciences, Australian National University: Canberra, Australia; 1994.
• [13]Mattnera J, Zawkoa G, Rossettoc M, Kraussb SL, Dixonb KW, Sivasithamparama K. Conservation genetics and implications for restoration of Hemigenia exilis (Lamiaceae), a serpentine endemic from Western Australia. Biol Conserv. 2002; 107:37-45.
• [14]Mekuria G, Collins G, Sedgley M. Genetic diversity within an isolated olive (Olea europaea L.) population in relation to feral spread. Sci Hort. 2002; 94:91-105.
• [15]Kovach W. MVSP-Multivariate Statistical Package (software) version 3.1. Kovach Computing Services, Pentraeth, UK; 1999.
• [16]Excoffier L, Smouse PE, Quattro JM. Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics. 1992; 131:479-491.
• [17]van Thiel PP, van Gool T, Faber WR, Leenstra T, Kager PA, Bart A. Variation in clinical presentation and genotype of causative Leishmania major strain in cutaneous leishmaniasis in north and south Afghanistan. Am J Trop Med Hyg. 2011; 85:60-63.
• [18]Garcia AL, Kindt A, Quispe-Tintaya KW, Bermudez H, Llanos A, Arevalo J et al.. American tegumentary leishmaniasis: antigen-gene polymorphism, taxonomy and clinical pleomorphism. Infect Genet Evol. 2005; 5:109-116.
• [19]Blackwell JM. Genetic susceptibility to leishmanial infections: studies in mice and man. Parasitology. 1996; 112 Suppl:S67-S74.
• [20]Reiner SL, Locksley RM. The regulation of immunity to Leishmania major. Annu Rev Immunol. 1995; 13:151-177.
• [21]Russo DM, Barral-Netto M, Barral A, Reed SG. Human T-cell responses in Leishmania infections. Prog Clin Parasitol. 1993; 3:119-144.
• [22]Mahmoudzadeh-Niknam H, Ajdary S, Riazi-Rad F, Mirzadegan E, Rezaeian A, Khaze V et al.. Molecular epidemiology of cutaneous leishmaniasis and heterogeneity of Leishmania major strains in Iran. Trop Med Int Health. 2012; 17:1335-1344.
• [23]Ghawar W, Toumi A, Snoussi MA, Chlif S, Zaatour A, Boukthir A et al.. Leishmania major infection among Psammomys obesus and Meriones shawi: reservoirs of zoonotic cutaneous leishmaniasis in Sidi Bouzid(central Tunisia). Vector Borne Zoonotic Dis. 2011; 11:1561-1568.
• [24]Fitzpatrick BM. Power and sample size for nested analysis of molecular variance. Mol Ecol. 2009; 18:3961-3966.
• [25]Al-Jawabreh A, Diezmann S, Muller M, Wirth T, Schnur LF, Strelkova MV et al.. Identification of geographically distributed sub-populations of Leishmania (Leishmania) major by microsatellite analysis. BMC Evol Biol. 2008; 8:183. BioMed Central Full Text
• [26]Seridi N, Amro A, Kuhls K, Belkaid M, Zidane C, Al-Jawabreh A et al.. Genetic polymorphism of Algerian Leishmania infantum strains revealed by multilocus microsatellite analysis. Microbes Infect. 2008; 10:1309-1315.
• [27]Mahnaz T, Al-Jawabreh A, Kuhls K, Schonian G. Multilocus microsatellite typing shows three different genetic clusters of Leishmania major in Iran. Microbes Infect. 2011; 13:937-942.
• [28]Hamarsheh O, Presber W, Yaghoobi-Ershadi MR, Amro A, Al-Jawabreh A, Sawalha S et al.. Population structure and geographical subdivision of the Leishmania major vector Phlebotomus papatasi as revealed by microsatellite variation. Med Vet Entomol. 2009; 23:69-77.
• [29]Elfari M, Schnur LF, Strelkova MV, Eisenberger CL, Jacobson RL, Greenblatt CL et al.. Genetic and biological diversity among populations of Leishmania major from Central Asia, the Middle East and Africa. Microbes Infect. 2005; 7:93-103.
• [30]Deperet C, Boinet E. Du bouton de Gafsa au camp de Sathonay. Archives Médicales et Pharmaceutiques Militaires. 1884; 3:296-302.
• [31]Jaouadi K, Depaquit J, Haouas N, Chaara D, Gorcii M, Chargui N et al.. Twenty-four new human cases of cutaneous leishmaniasis due to Leishmania killicki in Metlaoui, southwestern Tunisia. Probable role of Phlebotomus sergenti in the transmission. Acta Trop. 2012; 122:276-283.
• [32]Schonian G, Kuhls K, Mauricio IL. Molecular approaches for a better understanding of the epidemiology and population genetics of Leishmania. Parasitology. 2011; 138:405-425.