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

• Published in: PLoS neglected tropical diseases Vol. 13; no. 5; p. e0007403
• Main Authors: Kato, Hirotomo, Gomez, Eduardo A, Seki, Chisato, Furumoto, Hayato, Martini-Robles, Luiggi, Muzzio, Jenny, Calvopiña, Manuel, Velez, Lenin, Kubo, Makoto, Tabbabi, Ahmed, Yamamoto, Daisuke S, Hashiguchi, Yoshihisa
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.05.2019; Public Library of Science (PLoS)
• Subjects: Animals; Base Pair Mismatch; Biology and Life Sciences; Cell Nucleus - genetics; Cutaneous leishmaniasis; CytB gene; Cytochrome; Cytochrome b; Cytochrome P-450; Cytochromes; Discordance; Distribution; DNA; DNA, Kinetoplast; Ecuador; Enzymes; Epidemiology; Gene polymorphism; Genes; Genetic aspects; Genetic polymorphisms; Genetic research; Health aspects; Humans; Identification; Identification and classification; Infections; Leishmania; Leishmania - genetics; Leishmania - isolation & purification; Leishmania - physiology; Leishmaniasis; Leishmaniasis, Cutaneous - parasitology; Leishmaniasis, Cutaneous - transmission; Leishmaniasis, Mucocutaneous - parasitology; Leishmaniasis, Mucocutaneous - transmission; Medicine and Health Sciences; Meglumine antimoniate; Mitochondria - genetics; Mucocutaneous leishmaniasis; Nucleotide sequence; Parasites; Parasitic diseases; Parasitology; Pathogenicity; Pathogens; PCR; People and places; Polymerase Chain Reaction; Polymorphism; Polymorphism, Restriction Fragment Length; Protozoa; Protozoan Proteins - genetics; Psychodidae - parasitology; Psychodidae - physiology; Research and Analysis Methods; Restriction fragment length polymorphism; Signs and symptoms; Species; Strains; Symptoms; Tropical diseases; Vector-borne diseases; Vectors; Zoology; Ecuador; Japan
• ISSN: 1935-2735; 1935-2727; 1935-2735
• DOI: 10.1371/journal.pntd.0007403

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. 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. Although most results corresponded between the two analyses, PCR-RFLP analyses revealed distribution of hybrid strains between Leishmania (Viannia) guyanensis and L. (V.) braziliensis and between L. (V.) guyanensis and L. (V.) panamensis, of which the latter was firstly identified in Ecuador. Moreover, unexpected parasite strains having the kinetoplast cyt b gene of L. (V.) braziliensis and nuclear genes of L. (V.) guyanensis, L. (V.) panamensis, or a hybrid between L. (V.) guyanensis and L. (V.) panamensis were identified. This is the first report of the distribution of a protozoan parasite having mismatches between kinetoplast and nuclear genes, known as mito-nuclear discordance. The result demonstrated that genetically complex Leishmania strains are present in Ecuador. 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.