Detection of Leishmania tropica Using Nested-PCR and Some of Their Virulence Factors in Thi-Qar Province, Iraq

Main Article Content

Mohammed Hassan Flaih
Fadhil Abbas Al-Abady
Khwam Reissan Hussein

Abstract

Cutaneous leishmaniasis is one of endemic diseases in Iraq. It is considered as widely health problem and is an uncontrolled disease. The aim of the study is to identify of Leishmania species that cause skin lesions among patients in Thi-Qar Province, South of Iraq, also to detect some virulence factors of L. tropica. This study includes three local locations, Al-Hussein Teaching, Suq Al-Shyokh General and Al-Shatrah General Hospitals in Province for the period from the beginning of December 2018 to the end of September 2019. The samples were collected from 80 patients suffering from cutaneous leishmaniasis, both genders, different ages, various residence places and single and multiple lesions. Nested-PCR technique was used to amplify kinetoplast minicircle fragments DNA (kDNA). Conventional-PCR was performed for determination of some virulence factors (LPG1, GP63, CPA and PPG1 genes). The electrophoresis findings of kDNA gene showed two species of the parasite found in the study area, 65 samples were positive for cutaneous leishmaniasis, L. tropica at 750bp and L. major at 560bp. Generally, L. tropica (57.5%) was the most common specie and L. major (23.75%) appeared in a low level. There are no significant differences between the infections of males and females, while there are significant differences at the comparison between age groups. All virulence genes (LPG1, GP63, CPA and PPG1) appeared in all L. tropica isolates with high percentage (100%). L. tropica is the major specie which that caused CL in Thi-Qar province, while L. major appeared in low incidence. The virulence genes, which were reviewed, are necessary and important in pathogenesis of L. tropica.

Downloads

Download data is not yet available.

Article Details

How to Cite
1.
Flaih MH, Al-Abady FA, Hussein KR. Detection of Leishmania tropica Using Nested-PCR and Some of Their Virulence Factors in Thi-Qar Province, Iraq. Baghdad Sci.J [Internet]. 2021Mar.30 [cited 2021Apr.13];18(1(Suppl.):0700. Available from: https://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/4514
Section
article
Author Biographies

Mohammed Hassan Flaih, Medical Laboratory Department, Al-Nasiriya Technical Institute, Southern Technical University, Thi-Qar, Iraq.

تدريسي في قسم التقنيات المختبرية، المعهد التقني الناصرية، الجامعة التقنية الجنوبية، ذي قار، العراق

المرتبة العلمية مدرس

Fadhil Abbas Al-Abady, Department of Biology, College of Education for pure Sciences, University of Thi-Qar, Thi-Qar, Iraq.

تدريسي في قسم علوم الحياة، كلية التربية للعلوم الصرفة، جامعة ذي قار، ذي قار، العراق

المرتبة العلمي استاذ

Khwam Reissan Hussein, Medical Laboratory Department, Al-Nasiriya Technical Institute, Southern Technical University, Thi-Qar, Iraq

تدريسي في قسم التقنيات المختبرية، المعهد التقني الناصرية، الجامعة التقنية الجنوبية، ذي قار، العراق

المرتبة العلمية استاذ

References

Ghatee MA, Mirhendi H, Marashifard M, Kanannejad Z, Taylor WR, Sharifi I. Population Structure of Leishmania tropica Causing Anthroponotic Cutaneous Leishmaniasis in Southern Iran by PCR-RFLP of Kinetoplastid DNA. Biomed Res Int [Internet]. 2018;2018:1–11. Available from: https://doi.org/10.1155/2018/6049198

Aronson N, Herwaldt BL, Libman M, Pearson R, Lopez-velez R, Weina P, et al. Diagnosis and Treatment of Leishmaniasis : Clinical Practice Guidelines by the Infectious Diseases Society of America ( IDSA ) and the American Society of Tropical Medicine and Hygiene ( ASTMH ). Clin Infect Dis. 2016;63:202–64.

Sharma U, Singh S. Immunobiology of Leishmaniasis. Indian J Exp Biol. 2009;47(2009):412–23.

Alemayehu B, Alemayehu M. Leishmaniasis: A Review on Parasite, Vector and Reservoir Host. Heal Sci J. 2017;11(4):1–6.

Khosravi A, Sharifi I, Fekri A, Kermanizadeh A, Bamorovat M, Mostafavi M, et al. Clinical Features of Anthroponotic Cutaneous Leishmaniasis in a Major Focus, Southeastern Iran, 1994-2014. Iran J Parasitol. 2017;12(4):544–53.

Martínez-lópez M, Soto M, Iborra S, Sancho D. Leishmania Hijacks Myeloid Cells for Immune Escape. Front Microbiol. 2018;9:1–16.

Gupta G, Oghumu S, Satoskar AR. Mechanisms of Immune Evasion in Leishmaniasis Gaurva. Adv. Appl. Microbiol. 2013; 82:155-184. doi: 10.1016/B978-0-12-407679-2.00005-3.

Corrales RM, Sereno D, Mathieu-Daudé F. Deciphering the Leishmania exoproteome: What we know and what we can learn. FEMS Immunol Med Microbiol [Internet]. 2010;58(1):27–38. Available from: 10.1111/j.1574-695X.2009.00608.x

Atayde VD, Hassani K, Lira S, Borges R, Adhikari A, Martel C, et al. Leishmania Exosomes and other Virulence Factors : Impact on Innate Immune Response and Macrophage Functions. Cell Immunol [Internet]. 2016;1–35. Available from: http://dx.doi.org/10.1016/j.cellimm.2016.07.013

Forestier C, Gao Q, Boons G. Leishmania lipophosphoglycan : how to establish structure-activity relationships for this highly complex and multifunctional glycoconjugate ? Cellu and Infec Microbiol. 2015;4:1–7.

Singh KS. Human Emerging and Re-emerging Infections. 2nd ed. Singapore: Wiley; 2015. 1040 p.

Lázaro-Souza M, Matte C, Lima JB, Duque GA, Quintela-Carvalho G, Vivarini ÁC, et al. Leishmania infantum Lipophosphoglycan-Deficient Mutants: A Tool to Study Host Cell-Parasite Interplay. Front Microbiol [Internet]. 2018;9(626):1–10. Available from: doi: 10.3389/fmicb.2018.00626

Medina LS, Souza BA, Queiroz A, Guimarães LH, Machado PRL, Carvalho EM, et al. The gp63 Gene Cluster Is Highly Polymorphic in Natural Leishmania ( Viannia ) braziliensis Populations , but Functional Sites Are Conserved. PLoS One. 2016;1–13.

Yao C. Major Surface Protease of Trypanosomatids : One Size Fits All ? Infect Immun. 2010;78(1):22–31.

Hassani K, Shio MT, Martel C, Faubert D, Olivier M. Absence of Metalloprotease GP63 Alters the Protein Content of Leishmania Exosomes. PLoS One [Internet]. 2014;9(4):1–14. Available from: 10.1371/journal.pone.0095007

Oghumu S, Natarajan G, Satoskar AR. Pathogenesis of Leishmaniasis in Humans. Hum Emerg Re-emerging Infect. 2015;I:337–48.

Contreras I, Gomez M, Nguyen O, Shio M, McMaster R, Olivier M. Leishmania-induced inactivation of the macrophage transcription factor AP-1 is mediated by the parasite metalloprotease GP63. PLoS Pathog. 2010;6(10).

Rana S, Mahato JP, Kumar M, Sarsaiya S. Modeling and docking of Cysteine Protease-A (CPA) of Leishmania donovani. J Appl Pharm Sci. 2017;7(9):179–84.

Das P, Alam MN, Paik D, Karmakar K, De T, Chakraborti T. Protease Inhibitors in Potential Drug Development for Leishmaniasis. Indian J Biochem Biophys. 2013;50:363–76.

Siqueira-neto JL, Debnath A, Mccall L, Bernatchez JA, Ndao M, Reed SL, et al. Cysteine proteases in protozoan parasites. NEGLECTROPIC DISE [Internet]. 2018;12(8):1–20. Available from: https://doi.org/ 10.1371/journal.pntd.0006512

Mukhopadhyay S, Mandal C. Glycobiology of Leishmania donovani. Indian J Med Res. 2006;123:203–20.

Fernandez-prada C, Sharma M, Plourde M, Bresson E, Roy G, Leprohon P, et al. High-throughput Cos-Seq screen with intracellular Leishmania infantum for the discovery of novel drug-resistance mechanisms Christopher. Drugs Drug Resist [Internet]. 2018;8(2):165–73. Available from: https://doi.org/10.1016/j.ijpddr.2018.03.004

Aoki JI, Laranjeira-silva MF, Muxel SM, Floeter-winter LM. ScienceDirect The impact of arginase activity on virulence factors of Leishmania amazonensis. Curr Opin Microbiol [Internet]. 2019;52:110–5. Available from: https://doi.org/10.1016/j.mib.2019.06.003

Montgomery J, Curtis J, Handman E. Genetic and structural heterogeneity of proteophosphoglycans in Leishmania. Mol Biochem Parasitol. 2002;121:75–85.

Lima JB, Araújo-Santos T, Lázaro-Souza M, Carneiro AB, Ibraim IC, Jesus-Santos FH, et al. Leishmania infantum lipophosphoglycan induced- Prostaglandin E2 production in association with PPAR-γ expression via activation of Toll like receptors-1 and 2. Nature. 2017;7:1–11.

Mohammad FI, Hmood KA. Detection of Leishmania species by Nested-PCR and virulence factoes GIPLS, GP63 in L. Major by conventional-PCR. Biochem Cell Arch. 2018;18(2):2255–9.

Al-Difaie RS. Prevalence of Cutaneous Leishmaniasis in AL-Qadissia province and the evaluation of treatment response by pentostam with RT-PCR. Wasit University /College of Science; 2013.

Al-Hassani MKKT. Epidemiological, Molecular and Morphological Identification of cutaneous leishmaniasis and, It’s insect vectors in Eastern Al-Hamzah district,AlQadisiya province. Coll. Educat. AL-Qadisiya Univ.; 2016.

Izadi S, Mirhendi H, Jalalizand N, Khodadadi H, Mohebali M, Nekoeian S, et al. Molecular Epidemiological Survey of Cutaneous Leishmaniasis in Two Highly Endemic Metropolises of Iran , Application of FTA Cards for DNA Extraction From Giemsa-Stained Slides. Jundishapur J Microbiol. 2016;9(2):1–7.

Postigo JAR. Leishmaniasis in the world health organization eastern mediterranean region. Int J Antimicrob Agents [Internet]. 2010;36(1):62–5. Available from: doi: 10.1016/j.ijantimicag.2010.06.023

Abdolmajid F, Ghodratollah SS, Hushang R, Mojtaba MB, Ali MM, Abdolghayoum M. Identification of Leishmania species by kinetoplast DNA-polymerase chain reaction for the first time in Khaf district, Khorasan-e-Razavi province, Iran. Trop Parasitol. 2015;5(1):50–5.

Ramezany M, Sharifi I, Babaei Z, Ghasemi P, Almani N, Heshmatkhah A, et al. Geographical distribution and molecular characterization for cutaneous leishmaniasis species by sequencing and phylogenetic analyses of kDNA and ITS1 loci markers in south-eastern Iran. Pathog Glob Health [Internet]. 2018;112(3):132–41. Available from: https://doi.org/10.1080/20477724.2018.1447836

Azmi K, Nasereddin A, Ereqat S, Schnur L, Schonian G, Abdeen Z. Methods incorporating a polymerase chain reaction and restriction fragment length polymorphism and their use as a ‘ gold standard ’ in diagnosing Old World cutaneous leishmaniasis. Diagn Microbiol Infect Dis [Internet]. 2011;71(2):151–5. Available from: http://dx.doi.org/10.1016/j.diagmicrobio.2011.06.004

Cunze S, Kochmann J, Koch LK, Hasselmann KJQ, Klimpel S. Leishmaniasis in Eurasia and Africa : geographical distribution of vector species and pathogens. R Soc Open Sci [Internet]. 2019;6:1–12. Available from: http://dx.doi.org/10.1098/rsos.190334

Galgamuwa LS, Dharmaratne SD, Iddawela D. Leishmaniasis in Sri Lanka : spatial distribution and seasonal variations from 2009 to 2016. Parasit Vectors [Internet]. 2018;11(60):1–10. Available from: DOI 10.1186/s13071-018-2647-5

Al-bajalan MMM, Al-jaf SMA, Niranji SS, Abdulkareem DR, Al-kayali KK, Kato H. An outbreak of Leishmania major from an endemic to a non-endemic region posed a public health threat in Iraq from 2014-2017 : Epidemiological , molecular and phylogenetic studies. PLoS Negl Trop Dis. 2018;12(3):1–11.

Khosravi A, Sharifi I, Dortaj E, Afshar AA, Mostafavi M. The Present Status of Cutaneous Leishmaniasis in a Recently Emerged Focus in South-West of Kerman Province , Iran. Iran J Publ Heal. 2013;42(2):182–7.

El Hamouchi A, Daoui O, Kbaich MA, Mhaidi I, El Kacem S, Guizani I, et al. Epidemiological features of a recent zoonotic cutaneous leishmaniasis outbreak in Zagora province, southern Morocco. PLoS Negl Trop Dis. 2019;13(4):1–14.

Alsamarai AM, Alobaidi HS. Cutaneous leishmaniasis in Iraq. J Infect Dev Ctries. 2009;3(2):123–9.

Rahi AA. Cutaneous Leishmaniasis in Iraq: A clinicoepidemio-logical descriptive study. Sch J App Med Sci [Internet]. 2013;1(6):1021–5. Available from: http://saspublisher.com/wp-content/uploads/2013/12/SJAMS161021-1025.pdf

Hassan HF, Abbas SK, Shakoor DS. Epidemiological and hematological Investigation of Leishmania major. Kirkuk Unive J Sci. 2017;12(1):457–79.

Scala A, Micale N, Piperno A, Rescifina A, Schirmeister T, Kesselringc J, et al. Targeting of the Leishmania mexicana cysteine protease CPB2.8DCTE by decorated fused benzo[b] thiophene scaffol. R Soc Chem. 2016;6:30628–35.

Williams RA, Tetley L, Mottram JC, Coombs GH. Cysteine peptidases CPA and CPB are vital for autophagy and differentiation in Leishmania mexicana. Mol Microbiol [Internet]. 2006;61(3):655–74. Available from: doi:10.1111/j.1365-2958.2006.05274.x

Denise H, Poot J, Jiménez M, Ambit A, Herrmann DC, Vermeulen AN, et al. Studies on the CPA cysteine peptidase in the Leishmania infantum genome strain JPCM5. BioMed Cent. 2006;13:1–13.

Samant M, Sahasrabuddhe AA, Singh N, Gupta SK, Sundar S, Dube A. Proteophosphoglycan is differentially expressed in sodium stibogluconate-sensitive and resistant Indian clinical isolates of Leishmania donovani. Parasitol. 2007;134(9):1175–84.

Ilg T, Montgomery J, Stierhof Y, Handman E. Molecular Cloning and Characterization of a Novel Repeat-containing Leishmania major Gene , ppg1 , That Encodes a Membrane-associated Form of Proteophosphoglycan with a Putative Glycosylphosphatidylinositol Anchor. Biol Chem. 1999;274(44):31410–20.