The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have no conflicts of interest to declare. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the manuscript and its Supporting Information files.įunding: This study was financially supported by the Ministry of Education, Culture and Sports, Science and Technology (MEXT) of Japan (Grant Nos. Received: FebruAccepted: ApPublished: May 6, 2019Ĭopyright: © 2019 Kato et al. PLoS Negl Trop Dis 13(5):Įditor: Claudia Ida Brodskyn, Centro de Pesquisa Gonçalo Moniz-FIOCRUZ/BA, BRAZIL (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. Since hybrid strains of Leishmania were suggested to increase disease severity and may be transmitted by a wider range of sand fly species, careful epidemiological research, including clinical courses and vector research, will be needed.Ĭitation: Kato H, Gomez EA, Seki C, Furumoto H, Martini-Robles L, Muzzio J, et al. Although most results corresponded between the two analyses, PCR-RFLP revealed presence of unexpected genetically complex Leishmania strains having characteristics of hybrid and mito-nuclear discordance. In the present study, PCR-Restriction Fragment Length Polymorphism (RFLP) targeting multiple nuclear genes was employed, and the results were compared with those obtained by kinetoplast cytochrome b ( cyt b) gene sequence analysis, which is widely applied to species identification. The parasite species have been identified conventionally by multilocus enzyme electrophoresis (MLEE) and recently by genetic analysis such as sequencing and genotyping. Since infecting species are the major determinant of clinical outcomes, identification at the species level is important for the treatment and prognosis. Approximately 20 species are known to be pathogenic to humans, of which eight species have been recorded as causative agents of cutaneous and mucocutaneous leishmaniases in Ecuador. Leishmaniasis caused by intracellular protozoa of the genus Leishmania is a neglected tropical disease widely distributing worldwide, especially in tropical and subtropical areas. Since genetic exchanges such as hybrid formation were suggested to cause higher pathogenicity in Leishmania and may be transmitted by more species of sand flies, further country-wide epidemiological studies on clinical symptoms, as well as transmissible vectors, will be necessary. The result demonstrated that genetically complex Leishmania strains are present in Ecuador. This is the first report of the distribution of a protozoan parasite having mismatches between kinetoplast and nuclear genes, known as mito-nuclear discordance. Moreover, unexpected parasite strains having the kinetoplast cyt b gene of L. ) panamensis, of which the latter was firstly identified in Ecuador. Although most results corresponded between the two analyses, PCR-RFLP analyses revealed distribution of hybrid strains between Leishmania (Viannia) guyanensis and L. This method was applied to 92 clinical samples collected at 33 sites in 14 provinces of Ecuador, which have been identified at the species level by the kinetoplast cytochrome b ( cyt b) gene sequence analysis, and the results obtained by the two analyses were compared. PCR-Restriction Fragment Length Polymorphism (RFLP) analyses targeting multiple nuclear genes were established for the simple and practical identification of Leishmania species without using expensive equipment.
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