1921
Volume 88, Issue 3
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Abstract.

Leishmaniasis is a disease caused by different species belonging to the genus . It presents different epidemiological and clinical features and requires the development of rapid, sensitive techniques to improve specific diagnosis. In this study, we compared the traditional technique of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with PCR-fluorescent fragment length analysis (PCR-FFL). Fluorescently tagged primers, designed in the rRNA fragment ITS-1 and 7SL region, were used to amplify fragments, which were later digested and whose sizes were accurately determined using an automated DNA sequencer. We validated the technique using 19 strains from five cutaneous species before testing 36 clinical samples: 23 skin biopsies and 13 skin scrapings/lesion exudates on filter paper. In real diagnostic, PCR-FFL has proved to be quick, accurate, and more sensitive (83.3% testing the ITS-1 fragment and 94.4% testing the 7SL) than PCR-RFLP analysis (75% and 80.6%). Filter papers improved the specific diagnosis in both techniques using non-invasive samples.

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2013-03-06
2017-09-22
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References

  1. Desjeux P, , 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27: 305318.[Crossref]
  2. Ashford RW, , 2000. The leishmaniases as emerging and reemerging zoonoses. Int J Parasitol 30: 12691281.[Crossref]
  3. Pratlong F, Rioux JA, Marty P, Faraut-Gambarelli F, Dereure J, Lanotte G, Dedet JP, , 2004. Isoenzymatic analysis of 712 strains of Leishmania infantum in the south of France and relationship of enzymatic polymorphism to clinical and epidemiological features. J Clin Microbiol 42: 40774082.[Crossref]
  4. Bensoussan E, Nasereddin A, Jonas F, Schnur LF, Jaffe CL, , 2006. Comparison of PCR assays for diagnosis of cutaneous leishmaniasis. J Clin Microbiol 44: 14351439.[Crossref]
  5. Kuhls K, Mauricio IL, Pratlong F, Presber W, Schönian G, , 2005. Analysis of ribosomal DNA internal transcribed spacer sequences of the Leishmania donovani complex. Microbes Infect 7: 12241234.[Crossref]
  6. Schönian G, Kuhls K, Mauricio IL, , 2011. Molecular approaches for a better understanding of the epidemiology and population genetics of Leishmania . Parasitology 138: 405425.[Crossref]
  7. El Tai NO, El Fari M, Mauricio I, Miles MA, Oskam L, El Safi SH, Presber WH, Schönian G, , 2001. Leishmania donovani: intraspecific polymorphisms of Sudanese isolates revealed by PCR-based analyses and DNA sequencing. Exp Parasitol 97: 3544.[Crossref]
  8. Abda IB, 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: 1722.[Crossref]
  9. Azmi K, Nasereddin A, Ereqat S, Schönian G, Abdeen Z, , 2010. Identification of Old World Leishmania species by PCR-RFLP of the 7 spliced leader RNA gene and reverse dot blot assay. Trop Med Int Health 15: 872880.[Crossref]
  10. 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: 2024.[Crossref]
  11. Schönian G, Nasereddin A, Dinse N, Schweynoch C, Schallig HD, Presber W, Jaffe CL, , 2003. PCR diagnosis and characterization of Leishmania in local and imported clinical samples. Diagn Microbiol Infect Dis 47: 349358.[Crossref]
  12. Hamilton PB, Adams ER, Malele II, Gibson WC, , 2008. A novel, high-throughput technique for species identification reveals a new species of tsetse-transmitted trypanosome related to the Trypanosoma brucei subgenus, Trypanozoon . Infect Genet Evol 8: 2633.[Crossref]
  13. Adams E, Hamilton PB, Malele I, Gibson WC, , 2008. The identification, diversity and prevalence of trypanosomes in field caught tsetse in Tanzania using ITS-1 primers and fluorescent fragment length barcoding. Infect Genet Evol 8: 439444.[Crossref]
  14. Espiñeira M, Herrero B, Vieites JM, Santaclara FJ, , 2010. Detection and identification of anisakids in seafood by fragment length polymorphism analysis and PCR-RFLP of ITS-1 region. Food Contr 21: 10511060.[Crossref]
  15. Herrero B, Madriñán M, Vieites JM, Espiñeira M, , 2010. Rapid identification of seaweeds in food products by PCR combined with ALF-RFLP and FINS methodologies. J Agric Food Chem 58: 1158611592.[Crossref]
  16. Martín-Ezquerra G, Fisa R, Riera C, Rocamora V, Fernández-Casado A, Barranco C, Serra T, Baró T, Pujol RM, , 2009. Role of Leishmania spp. infestation in non-diagnostic cutaneous granulomatous lesions: report of a series of patients from a western Mediterranean area. Br J Dermatol 161: 320325.[Crossref]
  17. Zelazny AM, Fedorko DP, Li L, Neva FA, Fischer SH, , 2005. Evaluation of 7SL RNA gene sequences for the identification of Leishmania spp. Am J Trop Med Hyg 72: 415420.
  18. D'Amelio S, Mathiopoulos KD, Santos CP, Pugachev ON, Webb SC, Picanco M, , 2000. Genetic markers in ribosomal DNA for the identification of members of the genus Anisakis (Nematoda: Ascaridoidea) defined by polymerase chain reaction-based restriction fragment length polymorphism. Int J Parasitol 30: 223226.[Crossref]
  19. Umehara A, Kawakami Y, Matsumi T, Araki J, Uchida A, , 2006. Molecular identification of Anisakis simplex sensu stricto and Anisakis pegreffii (Nematoda: Anisakidae) from fish and cetacean in Japanese waters. Parasitol Int 55: 267271.[Crossref]
  20. Azmi K, Nasereddin A, Ereqat S, Schnur L, Schönian G, Abdeen Z, , 2011. 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 71: 151155.[Crossref]
  21. Nasereddin A, Jaffe CL, , 2010. Rapid diagnosis of Old World leishmaniasis by high-resolution melting analysis of the 7SL RNA gene. J Clin Microbiol 48: 22402242.[Crossref]
  22. Michaeli S, Podell D, Agabian N, Ullu E, , 1992. The 7SL RNA homologue of Trypanosoma brucei is closely related to mammalian 7SL RNA. Mol Biochem Parasitol 51: 5564.[Crossref]
  23. Ben-Shlomo H, Levitan A, Shay NE, Goncharov I, Michaeli S, , 1999. RNA editing associated with the generation of two distinct conformations of the trypanosomatid Leptomonas collosoma 7SL RNA. J Biol Chem 274: 2564225650.[Crossref]
  24. Meyer R, Hoffelein C, Candrian U, , 1995. Polymerase chain reaction restriction fragment length polymorphism analysis: a simple method for species identification in food. J AOAC Int 78: 15421551.
  25. Saitoh H, Ueda S, Kurosaki K, Kiuchi M, , 1998. The different mobility of complementary strands depends on the proportion AC/GT. Forensic Sci Int 91: 8190.[Crossref]
  26. Boggild AK, Ramos AP, Valencia BM, Veland N, Calderon F, Arevalo J, Low DE, Llanos-Cuentas A, , 2011. Diagnostic performance of filter paper lesion impression PCR for secondarily infected ulcers and nonulcerative lesions caused by cutaneous leishmaniasis. Am Soc Microbiol 49: 10971100.
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  • Received : 28 Jun 2012
  • Accepted : 09 Oct 2012

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