Authors:
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- ABSTRACT.
- INTRODUCTION
- MATERIALS AND METHODS
- RESULTS
- Polymerase chain reaction for detection of Leishmania DNA in clinical samples targeting the ITS1 genetic marker.
- ITS1-RFLP (HaeIII).
- Sequencing of ITS1 amplified products.
- Polymerase chain reaction for detection of Leishmania DNA in clinical samples targeting the hsp70 genetic marker.
- hsp70-RFLP (HaeIII).
- Sequencing of hsp70 amplified products.
- DISCUSSION
- CONCLUSION
- ACKNOWLEDGMENTS
- REFERENCES
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Ministério da Saúde, Secretaria de Vigilância em Saúde, Departamento de Vigilância Epidemiológica 2017. Manual de Vigilância e Leishmaniose Tegumentar Americana, 2nd ed. Brasília, Brazil: Ministério da Saúde.
-
2.
Lyra MR , Pimentel MIF , Madeira MF , Antonio LF , Lyra JPM , Fagundes A , Schubach AO , 2015. First report of cutaneous leishmaniasis caused by Leishmania (Leishmania) infantum chagasi in an urban area of Rio de Janeiro, Brazil. Rev Inst Med Trop São Paulo 57: 451–455.
-
3.
Ministério da Saúde, Secretaria de Vigilância em Saúde, Departamento de Vigilância Epidemiológica 2007. Manual de Vigilância e Leishmaniose Tegumentar Americana, 2nd ed. Brasília, Brazil: Ministério da Saúde.
-
4.
Ashford DA , Bozza M , Freire M , Miranda JC , Sherlock L , Eulálio C , Lopes U , Fernandes O , Degrave H , Badaro R , 1995. Comparison of the polymerase chain reaction on the detection of canine visceral leishmaniasis. Am J Trop Med Hyg 53: 251–255.
-
5.
Zampieri RA , Laranjeira-Silva MF , Muxel SM , Stocco de Lima AC , Shaw JJ , Floeter-Winter LM , 2016. High resolution melting analysis targeting hsp70 as a fast and efficient method for the discrimination of Leishmania species. PLoS Negl Trop Dis 10: e0004485.
-
6.
Schönian G , Naseriddin A , Dinse N , Schweynoch C , Schallig H , Presber W , Jaffe C , 2003. PCR diagnosis and characterization of Leishmania in local and imported clinical samples. Diagn Microbiol Infect Dis 47: 349–358.
-
7.
Marfurt J , Nasereddin A , Niederwieser I , Jaffe CJ , Beck HP , Felger I , 2003. Identification and differentiation of Leishmania species in clinical samples by PCR amplification of the miniexon sequence and subsequent restriction fragment length polymorphism analysis. J Clin Microbiol 41: 3147–3153.
-
8.
Asato Y , Oshiro M , Myint CK , Yamamoto Y , Kato H , Marco JD , Mimori T , Gomez EAL , Yoshihisa HY , Uezato H , 2009. Phylogenic analysis of the genus Leishmania by cytochrome b gene sequencing. Exp Parasitol 121: 352–361.
-
9.
Fraga J , Veland N , Montalvo AM , Praet N , Boggild AK , Valencia BM , Arévalo J , Llanos-Cuentas A , Dujardin JC , Van der Auwera G , 2012. Accurate and rapid species typing from cutaneous and mucocutaneous leishmaniasis lesions of the New World. Diagn Microbiol Infect Dis 74: 142–150.
-
10.
Silva LA , Sousa CS , Graça GC , Porrozzi R , Cupolillo E , 2010. Sequence analysis and PCR RFLP profiling of the hsp70 gene as a valuable tool for identifying Leishmania species associated with human leishmaniasis in Brazil. Infect Genet Evol 10: 77–83.
-
11.
Montalvo AM , Fraga J , Maes I , Dujardin JC , Van der Auwera G , 2012. Three new sensitive and specific heat shock protein 70 PCRs for global Leishmania species identification. Eur J Clin Microbiol Infect Dis 31: 1453–1461.
-
12.
Garcia L , Kindt A , Bermudez H , Llanos-Cuentas A , De Doncker S , Arevalo J , Wilber QTK , Dujardin JC , 2004. Culture-independent species typing of neotropical Leishmania for clinical validation of a PCR-based assay targeting heat shock protein 70 genes. J Clin Microbiol 42: 2294–2297.
-
13.
Khanra S , Datta S , Mondal D , Saha P , Bandopadhyay SK , Roy S , Manna M , 2012. RFLPs of ITS, ITS1 and hsp70 amplicons and sequencing of ITS1 of recent clinical isolates of Kala-azar from India and Bangladesh confirms the association of L. tropica with the disease. Acta Trop 124: 229–234.
-
14.
Mouttaki T , Morales-Yuste M , Merino-Espinosa G , Chiheb S , Fellah H , Martin-Sanchez J , Riyad M , 2014. Molecular diagnosis of cutaneous leishmaniasis and identification of the causative Leishmania species in Morocco by using three PCR-based assays. Parasit Vectors 7: 420.
-
15.
Montalvo Alvarez AM , Nodarse JF , Goodridge IM , Fidalgo LM , Marin M , Van Der Auwera G , Dujardin JC , Bernal ID , Muskus C , 2010. Differentiation of Leishmania (Viannia) panamensis and Leishmania (V.) guyanensis using BccI for hsp70 PCR-RFLP. Trans R Soc Trop Med Hyg 104: 364–367.
-
16.
Akhoundi M et al., 2017. Leishmania infections: molecular targets and diagnosis. Mol Aspects Med 57: 1–29.
-
17.
Gadisa E , Genetu A , Kuru T , Jirata D , Dagne K , Aseffa A , Gedamu L , 2007. Leishmania (Kinetoplastida): species typing with isoenzyme and PCR-RFLP from cutaneous leishmaniasis patients in Ethiopia. Exp Parasitol 115: 339–343.
-
18.
Montalvo AM , Fraga J , Monzote L , Montano I , De Doncker S , Dujardin JC , Van der Auwera G , 2010. Heatshock protein 70 PCR-RFLP: a universal simple tool for Leishmania species discrimination in the New and Old World. Parasitology 137: 1159–1168.
-
19.
Montalvo AM , Fraga J , El Safi S , Gramiccia M , Jaffe CL , Dujardin JC , Van der Auwera G , 2014. Direct Leishmania species typing in Old World clinical samples: evaluation of 3 sensitive methods based on the heat-shock protein 70 gene. Diagn Microbiol Infect Dis 80: 35–39.
-
20.
Ben Abdo I , de Monbrison F , Bousslimi N , Aoun K , Bouratbine A , Picot S , 2011. Advantages and limits of real time PCR assay and PCR-restriction fragment length polymorphism for the identification of cutaneous Leishmania species in Tunisia. Trans R Soc Trop Med Hyg 105: 17–22.
-
21.
Mosleh IM , Shonian G , Geith E , Al-Jawabreh A , Natsheh L , 2015. 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. Exp Parasitol 148: 81–85.
-
22.
Godoy NS , Vera DA , Regina MS , Okay TS , Braz LMA , 2018. Unusual clinical manifestations of Leishmania (L.) infantum chagasi in an HIV-coinfected patient and the relevance of ITS1-PCR-RFLP: a case report. Iran J Parasitol 13: 655–660.
-
23.
de Godoy NS , Lima-Junior MS , Lindoso JAL , Pereira-Chioccola VL , Okay TS , Braz LMA , 2020. A PCR and RFLP-based molecular diagnostic algorithm for visceral leishmaniasis. Asian Pac J Trop Med 13: 62–70.
-
24.
MacFarlane J , Blaxter ML , Bishop RP , Miles MA , Kelly JM , 1990. Identification and characterization of a Leishmania donovani antigen belonging to the 70 Kda heat shock protein family. Eur J Biochem 190: 377–384.
-
25.
Espada CR , Ortiz PA , Shaw JJ , Barral AMP , Costa JML , Uliana SRB , Coelho AC , 2018. Identification of Leishmania (Viannia) species and clinical isolates of Leishmania (Leishmania) amazonensis from Brazil using PCR-RFLP of the heat-shock protein 70 gene reveals some unexpected observations. Diagn Microbiol Infect Dis 91: 312–318.
-
26.
Ivancic-Jelecki J , Baricevic M , Santak M , Forcic D , 2006. Restriction enzyme cleavage of fluorescently labeled DNA fragments – analysis of the method and its usage in examination of digestion completeness. Anal Biochem 349: 277–284.
-
27.
Cupolillo E , Brahim LR , Toaldo CB , 2003. Genetic polymorphism and molecular epidemiology of Leishmania (Viannia) braziliensis from different hosts and geographic areas in Brazil. J Clin Microbiol 41: 3126–3132.
-
28.
Graça GC , Volpini AC , Romero GAS , Oliveira Neto MP , Hueb M , Porrozzi R , Boité MC , Cupolillo E , 2012. Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Mem Inst Oswaldo Cruz 107: 664–674.
-
29.
Navin TR , Arana BA , Arana FE , Berman JD , Chajon JF , 1992. Placebo-controlled clinical trial of sodium stibogluconate (Pentostam) versus ketoconazole for treating cutaneous leishmaniasis in Guatemala. J Infect Dis 165: 528–534.
-
30.
Romero GAS , Orge MGO , Guerra MVF , Paes MG , Macedo VO , Carvalho EM , 2005. Resposta de anticorpos em pacientes com leishmaniose cutânea infectados por Leishmania (Viannia) braziliensis ou Leishmania (Viannia) guyanensis no Brasil. Acta Trop 93: 49–56.
-
31.
Silva ACT , Cupolillo E , Volpini AC , Almeida R , Romero GA , 2006. Species diversity causing human cutaneous leishmaniasis in Rio Branco, state of Acre, Brazil. Trop Med Int Health 11: 1388–1398.
-
32.
Doni NY , Gurses G , Dikme R , Aksoy M , Yildiz Zeyrek F , Simsek Z , Satoskar AR , Varikuty S , Yesilova Y , 2020. Cutaneous leishmaniasis due to three Leishmania species among Syrian refugees in Sanliurfa, Southeastern Turkey. Acta Parasitol 65: 936–948.
-
33.
Evans TG , Krug EC , Wilson ME , Vasconcelos AW , Alencar JE , Pearson RD , 1989. Evaluation of antibody responses in American visceral leishmaniasis by ELISA and immunoblot. Mem Inst Oswaldo Cruz 84: 157–166.
-
34.
Ardehali S , Moattari A , Hatam GR , Hosseini SMH , Sharifi I , 2000. Characterization of Leishmania isolated in Iran: 1. Serotyping with species specific monoclonal antibodies. Acta Trop 75: 301–307.
-
35.
Reithinger R , Dujardin JC , 2007. Molecular diagnosis of leishmaniasis: current status and future applications. J Clin Microbiol 45: 21–25.
-
36.
Singh S , Sivakumar R , 2003. Recent advances in the diagnosis of leishmaniasis. J Postgrad Med 49: 55–60.
-
37.
Tavares CAP , Fernandes AP , Melo MN , 2003. Molecular diagnosis of leishmaniasis. Expert Rev Mol Diagn 3: 657–667.
-
38.
Vega-Lopez F , 2003. Diagnosis of cutaneous leishmaniasis. Curr Opin Infect Dis 16: 97–101.
-
39.
Kato H et al., 2019. PCR-RFLP analyses of Leishmania species causing cutaneous and mucocutaneous leishmaniasis revealed distribution of genetically complex strains with hybrid and mito-nuclear discordance in Ecuador. PLoS Negl Trop Dis 13: e0007403.
-
40.
Silva NL , de Silva VNH , Deerasinghe ATH , Rathnapala UL , Itoh M , Takagi H , Weerasooriya MV , Kato H , Yahathugoda TC , 2022. Development of a highly sensitive nested PCR and its application for the diagnosis of cutaneous leishmaniasis in Sri Lanka. Microorganisms 10: 990.
-
41.
Ertabaklar H , Ertug S , Çaliskan SO , Bozdogan B , 2016. Determination of Leishmania species by PCR-RFLP in the smear samples taken from the lesions of cutaneous leishmaniasis cases. Mikrobiyol Bul 50: 300–306.
-
42.
Hajjaran H , Vasigheh F , Mohebali M , Rezaei S , Mamishi S , Charedar S , 2011. Direct diagnosis of Leishmania species on serosity materials punctured from cutaneous leishmaniasis patients using PCR-RFLP. J Clin Lab Anal 25: 20–24.
-
43.
Hezari F , Niyyati M , Seyyed Tabaei SJ , Mohebali M , Moin Vaziri V , Behniafar H , Azargashb E , Taghipour N , 2016. Frequency of cutaneous leishmaniasis and species identification in suspected individuals from Golestan Province, Northern Iran in 2014. Iran J Public Health 45: 1348–1354.
-
44.
Kermanjani A , Akhlaghi L , Oormazdi H , Hadighi R , 2017. Isolation and identification of cutaneous leishmaniasis species by PCR-RFLP in Ilam province, the west of Iran. J Parasit Dis 41: 175–179.
-
45.
Lorestani RZ , Arjmand R , Saki J , Jelowdar A , Salmanzadeh S , 2020. Genotyping the causative agents of cutaneous leishmaniasis in Ahvaz southwest of Iran. J Parasit Dis 44: 436–440.
-
46.
Mohammadiha A , Dalimi A , Mahmoodi MR , Parian M , Pirestani M , Mohebali M , 2017. The PCR-RFLP-based detection and identification of the Leishmania species causing human cutaneous leishmaniasis in the Khorasan-Razavi Province, Northeast of Iran. J Arthropod Borne Dis 11: 383–392.
-
47.
Shaheen N , Qureshi NA , Fatima H , Afzal M , Alhewairini S , 2020. Molecular epidemiological survey of cutaneous leishmaniasis from Azad Jammu and Kashmir, Pakistan. Acta Trop 206: 105434.
-
48.
Bisetegn H , Zeleke AJ , Gadisa E , Shumie G , Damte D , Fenta T , Behaksra S , Bayih AG , 2020. Clinical, parasitological and molecular profiles of cutaneous leishmaniasis and its associated factors among clinically suspected patients attending Borumeda Hospital, North-East Ethiopia. PLoS Negl Trop Dis 14: e0008507.
-
49.
Lata S , Kumari S , Das R , Pasi S , Dhiman RC , 2021. Typical and atypical cutaneous leishmaniasis in Himachal Pradesh (India). Heliyon 7: e07282.
-
50.
Castillo-Ureta H , Zazueta-Moreno JM , Rendón-Maldonado JG , Torres-Avendaño JI , López-Moreno HS , Olimón-Andalón V , Salomón-Soto VM , Pérez-Sánchez FP , Torres-Montoya EH , 2019. First report of autochthonous canine leishmaniasis caused by Leishmania (L.) mexicana in Sinaloa, Mexico. Acta Trop 190: 253–256.
-
51.
Bilbao-Ramos P , Dea-Ayuela MA , Cardenas-Alegría O , Salamanca E , Santalla-Vargas JA , Benito C , Flores N , Bolás-Fernández F , 2017. Leishmaniasis in the major endemic region of Plurinational State of Bolivia: species identification, phylogeography and drug susceptibility implications. Acta Trop 176: 150–161.
-
52.
Almeida JV , de Souza CF , Fuzari AA , Joya CA , Valdivia HO , Bartholomeu DC , Brazil RP , 2021. Diagnosis and identification of Leishmania species in patients with cutaneous leishmaniasis in the state of Roraima, Brazil’s Amazon Region. Parasit Vectors 14: 32.
-
53.
Rigg CA , Calzada JE , Saldaña A , Perea M , Chaves LF , Valderrama A , 2019. Leishmania spp. infection rate and feeding patterns of sand flies (Diptera: Psychodidae) from a hyperendemic cutaneous leishmaniasis community in Panamá. Am J Trop Med Hyg 100: 798–807.
-
54.
Carvalho-Silva R , Ribeiro-da-Silva RC , Cruz LNPD , Oliveira MDS , Amoedo PM , Rebêlo JMM , Guimarães-E-Silva AS , Pinheiro VCS , 2022. Predominance of Leishmania (Leishmania) amazonensis DNA in Lutzomyia longipalpis sand flies (Diptera: Psychodidae) from an endemic area for leishmaniasis in Northeastern Brazil. Rev Inst Med Trop Sao Paulo 64: e32.
-
55.
Ávila MM , Brilhante AF , de Souza CF , Bevilacqua PD , Galati EAB , Brazil RP , 2018. Ecology, feeding and natural infection by Leishmania spp. of phlebotomine sand flies in an area of high incidence of American tegumentary leishmaniasis in the municipality of Rio Branco, Acre, Brazil. Parasit Vectors 11: 64.
-
56.
Muvdi-Arenas S , Ovalle-Bracho C , 2019. Leishmaniasis mucosa: una enfermedad olvidada, descripción e identificación de especies en 50 casos colombianos. Rev Inst Nac Salud Biom 39: 58–65.
-
57.
Miranda ADC et al., 2021. Molecular identification of parasites causing cutaneous leishmaniasis in Panama. Am J Trop Med Hyg 104: 1326–1334.
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Detection of Internal Transcribed Spacer 1 and hsp70 Genetic Markers Using Restriction Fragment Length Polymorphisms and Sequencing in Identification of Leishmania Species Causing Tegumentary Leishmaniasis in Brazil
Jaqueline Alves Delprete
Jaqueline Alves Delprete
Departamento de Doenças Infecciosas e Parasitárias, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil;
Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
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Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
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Livia Vieira de Almeida
Livia Vieira de Almeida
Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
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Alessandra Moraes Barros
Alessandra Moraes Barros
Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
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Rita de Cássia Soler
Rita de Cássia Soler
Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
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Amanda Azevedo Bittencourt
Amanda Azevedo Bittencourt
Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
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Expedito José de Albuquerque Luna
Expedito José de Albuquerque Luna
Departamento de Medicina Preventiva, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil;
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José Angelo Lauletta Lindoso
José Angelo Lauletta Lindoso
Departamento de Doenças Infecciosas e Parasitárias, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil;
Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
Laboratório de Protozoologia (LIM-56 HC-FMUSP), Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil
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Instituto de Infectologia Emílio Ribas, São Paulo, Brazil;
Laboratório de Protozoologia (LIM-56 HC-FMUSP), Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil
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Lúcia Maria Almeida Braz
Lúcia Maria Almeida Braz
Departamento de Medicina Preventiva, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil;
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ABSTRACT.
The identification of Leishmania species that cause tegumentary leishmaniasis (TL) is important for taxonomic and prognostic purposes. Molecular analysis using different Leishmania genomic targets is the most useful method for identifying Leishmania species. Therefore, we evaluated the performance of ribosomal RNA internal transcribed spacer 1 (ITS1) and heat shock protein (hsp70) genetic markers by polymerase chain reaction (PCR), followed by restriction fragment length polymorphism analysis (RFLP) and sequencing, for identification of Leishmania species. Samples from 84 Brazilian patients were amplified. Internal transcribed spacer 1 PCR followed by RFLP (HaeIII) [ITS1-RFLP (HaeIII)] identified 46.4% (39/84) of the samples as compatible with the Viannia subgenus. Internal transcribed spacer 1 PCR followed by sequencing (ITS1-sequencing) identified Leishmania (Viannia) braziliensis in 91.7% (77/84) of the TL samples, Leishmania (Leishmania) amazonensis in 3.6% (3/84), L. (V.) guyanensis in 2.4% (2/84), and L. (L.) infantum in 1.2% (1/84). One of the samples showed the same proportion of similarity with L. (V.) guyanensis and L. (V.) panamensis. hsp70 nested PCR followed by RFLP (HaeIII) [nested hsp70-RFLP (HaeIII)] identified 91.7% (77/84) of the samples as compatible with L. (V.) braziliensis/L. (V.) naiffi, 3.6% (3/84) with L. (L.) amazonensis, 1.2% (1/84) with L. (L.) infantum, and 3.6% (3/84) with L. (V.) guyanensis. hsp70 PCR followed by sequencing (hsp70-sequencing) identified L. (V.) braziliensis in 91.7% (77/84) of the TL samples, L. (L.) amazonensis in 3.6% (3/84), L. (V.) guyanensis in 3.6% (3/84), and L. (L.) infantum in 1.2% (1/84). Our findings clearly showed that nested hsp70-RFLP (HaeIII) is better than ITS1-RFLP (HaeIII) and that ITS1 or hsp70 PCR followed by sequencing was adequate for identifying Leishmania species. We also found that Leishmania (Viannia) braziliensis is the most common species causing TL in Brazil. Therefore, sequencing multiple target genes such as ITS1 and hsp 70 is more accurate than RFLP for identifying Leishmania species.
Author Notes
Financial support: This study was funded by grant
Authors’ addresses: Jaqueline Alves Delprete, Departamento de Doenças Infecciosas e Parasitárias, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil, and Instituto de Infectologia Emílio Ribas, São Paulo, Brazil, E-mail: delprete@usp.br. Livia Vieira de Almeida, Alessandra Moraes Barros, Rita de Cássia Soler, and Amanda Azevedo Bittencourt, Instituto de Infectologia Emílio Ribas, São Paulo, Brazil, E-mails: livia.almeida@emilioribas.sp.gov.br, alebarros.derma@gmail.com, rita.soler@emilioribas.sp.gov.br, and am.bitten@gmail.com. Expedito José de Albuquerque Luna and Lúcia Maria Almeida Braz, Departamento de Medicina Preventiva, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil, E-mails: eluna@usp.br and lmabraz@usp.br. José Angelo Lauletta Lindoso, Departamento de Doenças Infecciosas e Parasitárias, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brazil, Instituto de Infectologia Emílio Ribas, São Paulo, Brazil, and Laboratório de Protozoologia (LIM-56 HC-FMUSP), Instituto de Medicina Tropical, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil, E-mail: jlindoso@usp.br.
- ABSTRACT.
- INTRODUCTION
- MATERIALS AND METHODS
- RESULTS
- Polymerase chain reaction for detection of Leishmania DNA in clinical samples targeting the ITS1 genetic marker.
- ITS1-RFLP (HaeIII).
- Sequencing of ITS1 amplified products.
- Polymerase chain reaction for detection of Leishmania DNA in clinical samples targeting the hsp70 genetic marker.
- hsp70-RFLP (HaeIII).
- Sequencing of hsp70 amplified products.
- DISCUSSION
- CONCLUSION
- ACKNOWLEDGMENTS
- REFERENCES
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Ministério da Saúde, Secretaria de Vigilância em Saúde, Departamento de Vigilância Epidemiológica 2017. Manual de Vigilância e Leishmaniose Tegumentar Americana, 2nd ed. Brasília, Brazil: Ministério da Saúde.
-
2.
Lyra MR , Pimentel MIF , Madeira MF , Antonio LF , Lyra JPM , Fagundes A , Schubach AO , 2015. First report of cutaneous leishmaniasis caused by Leishmania (Leishmania) infantum chagasi in an urban area of Rio de Janeiro, Brazil. Rev Inst Med Trop São Paulo 57: 451–455.
-
3.
Ministério da Saúde, Secretaria de Vigilância em Saúde, Departamento de Vigilância Epidemiológica 2007. Manual de Vigilância e Leishmaniose Tegumentar Americana, 2nd ed. Brasília, Brazil: Ministério da Saúde.
-
4.
Ashford DA , Bozza M , Freire M , Miranda JC , Sherlock L , Eulálio C , Lopes U , Fernandes O , Degrave H , Badaro R , 1995. Comparison of the polymerase chain reaction on the detection of canine visceral leishmaniasis. Am J Trop Med Hyg 53: 251–255.
-
5.
Zampieri RA , Laranjeira-Silva MF , Muxel SM , Stocco de Lima AC , Shaw JJ , Floeter-Winter LM , 2016. High resolution melting analysis targeting hsp70 as a fast and efficient method for the discrimination of Leishmania species. PLoS Negl Trop Dis 10: e0004485.
-
6.
Schönian G , Naseriddin A , Dinse N , Schweynoch C , Schallig H , Presber W , Jaffe C , 2003. PCR diagnosis and characterization of Leishmania in local and imported clinical samples. Diagn Microbiol Infect Dis 47: 349–358.
-
7.
Marfurt J , Nasereddin A , Niederwieser I , Jaffe CJ , Beck HP , Felger I , 2003. Identification and differentiation of Leishmania species in clinical samples by PCR amplification of the miniexon sequence and subsequent restriction fragment length polymorphism analysis. J Clin Microbiol 41: 3147–3153.
-
8.
Asato Y , Oshiro M , Myint CK , Yamamoto Y , Kato H , Marco JD , Mimori T , Gomez EAL , Yoshihisa HY , Uezato H , 2009. Phylogenic analysis of the genus Leishmania by cytochrome b gene sequencing. Exp Parasitol 121: 352–361.
-
9.
Fraga J , Veland N , Montalvo AM , Praet N , Boggild AK , Valencia BM , Arévalo J , Llanos-Cuentas A , Dujardin JC , Van der Auwera G , 2012. Accurate and rapid species typing from cutaneous and mucocutaneous leishmaniasis lesions of the New World. Diagn Microbiol Infect Dis 74: 142–150.
-
10.
Silva LA , Sousa CS , Graça GC , Porrozzi R , Cupolillo E , 2010. Sequence analysis and PCR RFLP profiling of the hsp70 gene as a valuable tool for identifying Leishmania species associated with human leishmaniasis in Brazil. Infect Genet Evol 10: 77–83.
-
11.
Montalvo AM , Fraga J , Maes I , Dujardin JC , Van der Auwera G , 2012. Three new sensitive and specific heat shock protein 70 PCRs for global Leishmania species identification. Eur J Clin Microbiol Infect Dis 31: 1453–1461.
-
12.
Garcia L , Kindt A , Bermudez H , Llanos-Cuentas A , De Doncker S , Arevalo J , Wilber QTK , Dujardin JC , 2004. Culture-independent species typing of neotropical Leishmania for clinical validation of a PCR-based assay targeting heat shock protein 70 genes. J Clin Microbiol 42: 2294–2297.
-
13.
Khanra S , Datta S , Mondal D , Saha P , Bandopadhyay SK , Roy S , Manna M , 2012. RFLPs of ITS, ITS1 and hsp70 amplicons and sequencing of ITS1 of recent clinical isolates of Kala-azar from India and Bangladesh confirms the association of L. tropica with the disease. Acta Trop 124: 229–234.
-
14.
Mouttaki T , Morales-Yuste M , Merino-Espinosa G , Chiheb S , Fellah H , Martin-Sanchez J , Riyad M , 2014. Molecular diagnosis of cutaneous leishmaniasis and identification of the causative Leishmania species in Morocco by using three PCR-based assays. Parasit Vectors 7: 420.
-
15.
Montalvo Alvarez AM , Nodarse JF , Goodridge IM , Fidalgo LM , Marin M , Van Der Auwera G , Dujardin JC , Bernal ID , Muskus C , 2010. Differentiation of Leishmania (Viannia) panamensis and Leishmania (V.) guyanensis using BccI for hsp70 PCR-RFLP. Trans R Soc Trop Med Hyg 104: 364–367.
-
16.
Akhoundi M et al., 2017. Leishmania infections: molecular targets and diagnosis. Mol Aspects Med 57: 1–29.
-
17.
Gadisa E , Genetu A , Kuru T , Jirata D , Dagne K , Aseffa A , Gedamu L , 2007. Leishmania (Kinetoplastida): species typing with isoenzyme and PCR-RFLP from cutaneous leishmaniasis patients in Ethiopia. Exp Parasitol 115: 339–343.
-
18.
Montalvo AM , Fraga J , Monzote L , Montano I , De Doncker S , Dujardin JC , Van der Auwera G , 2010. Heatshock protein 70 PCR-RFLP: a universal simple tool for Leishmania species discrimination in the New and Old World. Parasitology 137: 1159–1168.
-
19.
Montalvo AM , Fraga J , El Safi S , Gramiccia M , Jaffe CL , Dujardin JC , Van der Auwera G , 2014. Direct Leishmania species typing in Old World clinical samples: evaluation of 3 sensitive methods based on the heat-shock protein 70 gene. Diagn Microbiol Infect Dis 80: 35–39.
-
20.
Ben Abdo I , de Monbrison F , Bousslimi N , Aoun K , Bouratbine A , Picot S , 2011. Advantages and limits of real time PCR assay and PCR-restriction fragment length polymorphism for the identification of cutaneous Leishmania species in Tunisia. Trans R Soc Trop Med Hyg 105: 17–22.
-
21.
Mosleh IM , Shonian G , Geith E , Al-Jawabreh A , Natsheh L , 2015. 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. Exp Parasitol 148: 81–85.
-
22.
Godoy NS , Vera DA , Regina MS , Okay TS , Braz LMA , 2018. Unusual clinical manifestations of Leishmania (L.) infantum chagasi in an HIV-coinfected patient and the relevance of ITS1-PCR-RFLP: a case report. Iran J Parasitol 13: 655–660.
-
23.
de Godoy NS , Lima-Junior MS , Lindoso JAL , Pereira-Chioccola VL , Okay TS , Braz LMA , 2020. A PCR and RFLP-based molecular diagnostic algorithm for visceral leishmaniasis. Asian Pac J Trop Med 13: 62–70.
-
24.
MacFarlane J , Blaxter ML , Bishop RP , Miles MA , Kelly JM , 1990. Identification and characterization of a Leishmania donovani antigen belonging to the 70 Kda heat shock protein family. Eur J Biochem 190: 377–384.
-
25.
Espada CR , Ortiz PA , Shaw JJ , Barral AMP , Costa JML , Uliana SRB , Coelho AC , 2018. Identification of Leishmania (Viannia) species and clinical isolates of Leishmania (Leishmania) amazonensis from Brazil using PCR-RFLP of the heat-shock protein 70 gene reveals some unexpected observations. Diagn Microbiol Infect Dis 91: 312–318.
-
26.
Ivancic-Jelecki J , Baricevic M , Santak M , Forcic D , 2006. Restriction enzyme cleavage of fluorescently labeled DNA fragments – analysis of the method and its usage in examination of digestion completeness. Anal Biochem 349: 277–284.
-
27.
Cupolillo E , Brahim LR , Toaldo CB , 2003. Genetic polymorphism and molecular epidemiology of Leishmania (Viannia) braziliensis from different hosts and geographic areas in Brazil. J Clin Microbiol 41: 3126–3132.
-
28.
Graça GC , Volpini AC , Romero GAS , Oliveira Neto MP , Hueb M , Porrozzi R , Boité MC , Cupolillo E , 2012. Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species. Mem Inst Oswaldo Cruz 107: 664–674.
-
29.
Navin TR , Arana BA , Arana FE , Berman JD , Chajon JF , 1992. Placebo-controlled clinical trial of sodium stibogluconate (Pentostam) versus ketoconazole for treating cutaneous leishmaniasis in Guatemala. J Infect Dis 165: 528–534.
-
30.
Romero GAS , Orge MGO , Guerra MVF , Paes MG , Macedo VO , Carvalho EM , 2005. Resposta de anticorpos em pacientes com leishmaniose cutânea infectados por Leishmania (Viannia) braziliensis ou Leishmania (Viannia) guyanensis no Brasil. Acta Trop 93: 49–56.
-
31.
Silva ACT , Cupolillo E , Volpini AC , Almeida R , Romero GA , 2006. Species diversity causing human cutaneous leishmaniasis in Rio Branco, state of Acre, Brazil. Trop Med Int Health 11: 1388–1398.
-
32.
Doni NY , Gurses G , Dikme R , Aksoy M , Yildiz Zeyrek F , Simsek Z , Satoskar AR , Varikuty S , Yesilova Y , 2020. Cutaneous leishmaniasis due to three Leishmania species among Syrian refugees in Sanliurfa, Southeastern Turkey. Acta Parasitol 65: 936–948.
-
33.
Evans TG , Krug EC , Wilson ME , Vasconcelos AW , Alencar JE , Pearson RD , 1989. Evaluation of antibody responses in American visceral leishmaniasis by ELISA and immunoblot. Mem Inst Oswaldo Cruz 84: 157–166.
-
34.
Ardehali S , Moattari A , Hatam GR , Hosseini SMH , Sharifi I , 2000. Characterization of Leishmania isolated in Iran: 1. Serotyping with species specific monoclonal antibodies. Acta Trop 75: 301–307.
-
35.
Reithinger R , Dujardin JC , 2007. Molecular diagnosis of leishmaniasis: current status and future applications. J Clin Microbiol 45: 21–25.
-
36.
Singh S , Sivakumar R , 2003. Recent advances in the diagnosis of leishmaniasis. J Postgrad Med 49: 55–60.
-
37.
Tavares CAP , Fernandes AP , Melo MN , 2003. Molecular diagnosis of leishmaniasis. Expert Rev Mol Diagn 3: 657–667.
-
38.
Vega-Lopez F , 2003. Diagnosis of cutaneous leishmaniasis. Curr Opin Infect Dis 16: 97–101.
-
39.
Kato H et al., 2019. PCR-RFLP analyses of Leishmania species causing cutaneous and mucocutaneous leishmaniasis revealed distribution of genetically complex strains with hybrid and mito-nuclear discordance in Ecuador. PLoS Negl Trop Dis 13: e0007403.
-
40.
Silva NL , de Silva VNH , Deerasinghe ATH , Rathnapala UL , Itoh M , Takagi H , Weerasooriya MV , Kato H , Yahathugoda TC , 2022. Development of a highly sensitive nested PCR and its application for the diagnosis of cutaneous leishmaniasis in Sri Lanka. Microorganisms 10: 990.
-
41.
Ertabaklar H , Ertug S , Çaliskan SO , Bozdogan B , 2016. Determination of Leishmania species by PCR-RFLP in the smear samples taken from the lesions of cutaneous leishmaniasis cases. Mikrobiyol Bul 50: 300–306.
-
42.
Hajjaran H , Vasigheh F , Mohebali M , Rezaei S , Mamishi S , Charedar S , 2011. Direct diagnosis of Leishmania species on serosity materials punctured from cutaneous leishmaniasis patients using PCR-RFLP. J Clin Lab Anal 25: 20–24.
-
43.
Hezari F , Niyyati M , Seyyed Tabaei SJ , Mohebali M , Moin Vaziri V , Behniafar H , Azargashb E , Taghipour N , 2016. Frequency of cutaneous leishmaniasis and species identification in suspected individuals from Golestan Province, Northern Iran in 2014. Iran J Public Health 45: 1348–1354.
-
44.
Kermanjani A , Akhlaghi L , Oormazdi H , Hadighi R , 2017. Isolation and identification of cutaneous leishmaniasis species by PCR-RFLP in Ilam province, the west of Iran. J Parasit Dis 41: 175–179.
-
45.
Lorestani RZ , Arjmand R , Saki J , Jelowdar A , Salmanzadeh S , 2020. Genotyping the causative agents of cutaneous leishmaniasis in Ahvaz southwest of Iran. J Parasit Dis 44: 436–440.
-
46.
Mohammadiha A , Dalimi A , Mahmoodi MR , Parian M , Pirestani M , Mohebali M , 2017. The PCR-RFLP-based detection and identification of the Leishmania species causing human cutaneous leishmaniasis in the Khorasan-Razavi Province, Northeast of Iran. J Arthropod Borne Dis 11: 383–392.
-
47.
Shaheen N , Qureshi NA , Fatima H , Afzal M , Alhewairini S , 2020. Molecular epidemiological survey of cutaneous leishmaniasis from Azad Jammu and Kashmir, Pakistan. Acta Trop 206: 105434.
-
48.
Bisetegn H , Zeleke AJ , Gadisa E , Shumie G , Damte D , Fenta T , Behaksra S , Bayih AG , 2020. Clinical, parasitological and molecular profiles of cutaneous leishmaniasis and its associated factors among clinically suspected patients attending Borumeda Hospital, North-East Ethiopia. PLoS Negl Trop Dis 14: e0008507.
-
49.
Lata S , Kumari S , Das R , Pasi S , Dhiman RC , 2021. Typical and atypical cutaneous leishmaniasis in Himachal Pradesh (India). Heliyon 7: e07282.
-
50.
Castillo-Ureta H , Zazueta-Moreno JM , Rendón-Maldonado JG , Torres-Avendaño JI , López-Moreno HS , Olimón-Andalón V , Salomón-Soto VM , Pérez-Sánchez FP , Torres-Montoya EH , 2019. First report of autochthonous canine leishmaniasis caused by Leishmania (L.) mexicana in Sinaloa, Mexico. Acta Trop 190: 253–256.
-
51.
Bilbao-Ramos P , Dea-Ayuela MA , Cardenas-Alegría O , Salamanca E , Santalla-Vargas JA , Benito C , Flores N , Bolás-Fernández F , 2017. Leishmaniasis in the major endemic region of Plurinational State of Bolivia: species identification, phylogeography and drug susceptibility implications. Acta Trop 176: 150–161.
-
52.
Almeida JV , de Souza CF , Fuzari AA , Joya CA , Valdivia HO , Bartholomeu DC , Brazil RP , 2021. Diagnosis and identification of Leishmania species in patients with cutaneous leishmaniasis in the state of Roraima, Brazil’s Amazon Region. Parasit Vectors 14: 32.
-
53.
Rigg CA , Calzada JE , Saldaña A , Perea M , Chaves LF , Valderrama A , 2019. Leishmania spp. infection rate and feeding patterns of sand flies (Diptera: Psychodidae) from a hyperendemic cutaneous leishmaniasis community in Panamá. Am J Trop Med Hyg 100: 798–807.
-
54.
Carvalho-Silva R , Ribeiro-da-Silva RC , Cruz LNPD , Oliveira MDS , Amoedo PM , Rebêlo JMM , Guimarães-E-Silva AS , Pinheiro VCS , 2022. Predominance of Leishmania (Leishmania) amazonensis DNA in Lutzomyia longipalpis sand flies (Diptera: Psychodidae) from an endemic area for leishmaniasis in Northeastern Brazil. Rev Inst Med Trop Sao Paulo 64: e32.
-
55.
Ávila MM , Brilhante AF , de Souza CF , Bevilacqua PD , Galati EAB , Brazil RP , 2018. Ecology, feeding and natural infection by Leishmania spp. of phlebotomine sand flies in an area of high incidence of American tegumentary leishmaniasis in the municipality of Rio Branco, Acre, Brazil. Parasit Vectors 11: 64.
-
56.
Muvdi-Arenas S , Ovalle-Bracho C , 2019. Leishmaniasis mucosa: una enfermedad olvidada, descripción e identificación de especies en 50 casos colombianos. Rev Inst Nac Salud Biom 39: 58–65.
-
57.
Miranda ADC et al., 2021. Molecular identification of parasites causing cutaneous leishmaniasis in Panama. Am J Trop Med Hyg 104: 1326–1334.
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