Intraspecific genetic variability in a population of Moroccan Leishmania infantum revealed by PCR-RFLP of kDNA minicircles

Hamouchi, Adil El; Ejghal, Rajaa; Hida, M.; Lemrani, Meryem

doi:10.1016/j.actatropica.2017.02.016

Cited by 13 publications

(34 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…kDNA sequence variability was also investigated by in silico RFLP analysis with the RestrictionMapper online program (://www.restrictionmapper.org/). A panel of nine endonucleases ( Bgl II, Bme1390 I, Dde I, Hpa II, Rsa I, Vsp I, Pst I, Sfc I, Xap I) was used, and according to the number and size of the fragments obtained, sequences were classified into the RFLP genotypes described by Cortes et al () and El Hamouchi et al ().…”

Section: Methodsmentioning

confidence: 99%

“…restr ictio nmapp er.org/). A panel of nine endonucleases (BglII, Bme1390I, DdeI, HpaII, RsaI, VspI, PstI, SfcI, XapI) was used, and according to the number and size of the fragments obtained, sequences were classified into the RFLP genotypes described by Cortes et al (2006) andEl Hamouchi et al (2017).…”

Section: Restriction Fragment Length Polymorphism (Rflp) Analysismentioning

confidence: 99%

“…Restriction fragment length polymorphism preceded by polymerase chain reaction (PCR-RFLP) on various gene targets, including kDNA, has been widely used for Leishmania typing (Chicharro et al, 2002;Cortes et al, 2006;Salvatore et al, 2016;Sobrino et al, 2008), and to date, 18 different RFLP genotypes named from A to R have been characterized by this method (Cortes et al, 2006;da Silva et al, 2015;El Hamouchi, Ejghal, Hida, & Lemrani, 2017). The above genotypes have been also related with the human patients' immunological status, with genotypes A and B being more commonly found in immunocompromised and immunocompetent individuals, respectively (Cortes et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Ortuño

Latrofa

Iborra

et al. 2019

Zoonoses and Public Health

View full text Add to dashboard Cite

Leishmania infantum causes human and canine leishmaniosis. The parasite, transmitted by phlebotomine sand flies, infects species other than dogs and people, including wildlife, although their role as reservoirs of infection remains unknown for most species. Molecular typing of parasites to investigate genetic variability and evolutionary proximity can help understand transmission cycles and designing control strategies. We investigated Leishmania DNA variability in kinetoplast (kDNA) and internal transcribed spacer 2 (ITS2) sequences in asymptomatically infected wildlife (n = 58) and symptomatically and asymptomatically infected humans (n = 38) and dogs (n = 15) from south‐east Spain, using single nucleotide polymorphisms (SNPs) and in silico restriction fragment length polymorphism (RFLP) analyses. All ITS2 sequences (n = 76) displayed a 99%–100% nucleotide identity with a L. infantum reference sequence, except one with a 98% identity to a reference Leishmania panamensis sequence, from an Ecuadorian patient. No heterogeneity was recorded in the 73 L. infantum ITS2 sequences except for one SNP in a human parasite sequence. In contrast, kDNA analysis of 44 L. infantum sequences revealed 11 SNP genotypes (nucleotide variability up to 4.3%) and four RFLP genotypes including B, F and newly described S and T genotypes. Genotype frequency was significantly greater in symptomatic compared to asymptomatic individuals. Both methods similarly grouped parasites as predominantly or exclusively found in humans, in dogs, in wildlife or in all three of them. Accordingly, the phylogenetic analysis of kDNA sequences revealed three main clusters, two as a paraphyletic human parasites clade and a third including dogs, people and wildlife parasites. Results suggest that Leishmania infantum genetics is complex even in small geographical areas and that, probably, several independent transmission cycles take place simultaneously including some connecting animals and humans. Investigating these transmission networks may be useful in understanding the transmission dynamics, infection risk and therefore in planning L. infantum control strategies.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Restriction Fragment Length Polymorphism (Rflp) Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Ortuño

Latrofa

Iborra

et al. 2019

Zoonoses and Public Health

View full text Add to dashboard Cite

show abstract

“…Different PCR-based methods have been employed to ascertain the intraspecific genetic variability of Leishmania and have contributed to the development of epidemiological studies [ 16 , 17 ]. PCR amplification of kinetoplast DNA (kDNA) minicircles followed by analysis of restriction fragment length polymorphism (RFLP) has shown the genetic diversity between closely related strains of L. infantum MON-1 [ 18 – 20 ]. Further genetic diversity within Leishmania strains has been elucidated by random amplification of polymorphic DNA (RAPD) [ 21 , 22 ], PCR-RFLP of the antigen-encoding genes gp 63 and cpb [ 23 , 24 ], sequence analysis of intergenic spacer regions (ITS) [ 25 , 26 ], multilocus microsatellite typing (MLMT) [ 18 , 27 – 29 ], and multilocus sequence typing (MLST) [ 30 – 32 ].…”

Section: Introductionmentioning

confidence: 99%

Genetic polymorphism in Leishmania infantum isolates from human and animals determined by nagt PCR-RFLP

et al. 2018

Self Cite

View full text Add to dashboard Cite

BackgroundLeishmania infantum is the causative agent of human visceral leishmaniasis (VL) and sporadic human cutaneous leishmaniasis (CL) in the Mediterranean region. The genetic variation of the Leishmania parasites may result in different phenotypes that can be associated with the geographical distribution and diversity of the clinical manifestations. The main objective of this study was to explore the genetic polymorphism in L. infantum isolates from human and animal hosts in different regions of Morocco.MethodsThe intraspecific genetic variability of 40 Moroccan L. infantum MON-1 strains isolated from patients with VL (n = 31) and CL (n = 2) and from dogs (n = 7) was evaluated by PCR-RFLP of nagt, a single-copy gene encoding N-acetylglucosamine-1-phosphate transferase. For a more complete analysis of L. infantum polymorphism, we included the restriction patterns of nagt from 17 strains available in the literature and patterns determined by in-silico digestion of three sequences from the GenBank database.ResultsMoroccan L. infantum strains presented a certain level of genetic diversity and six distinct nagt-RFLP genotypes were identified. Three of the six genotypes were exclusively identified in the Moroccan population of L. infantum: variant M1 (15%), variant M2 (7.5%), and variant M3 (2.5%). The most common genotype (65%), variant 2 (2.5%), and variant 4 (7.5%), were previously described in several countries with endemic leishmaniasis. Phylogenetic analysis segregated our L. infantum population into two distinct clusters, whereas variant M2 was clearly distinguished from both cluster I and cluster II. This distribution highlights the degree of genetic variability among the Moroccan L. infantum population.ConclusionThe nagt PCR-RFLP method presented here showed an important genetic heterogeneity among Moroccan L. infantum strains isolated from human and canine reservoirs with 6 genotypes identified. Three of the six Moroccan nagt genotypes, have not been previously described and support the particular genetic diversity of the Moroccan L. infantum population reported in other studies.Electronic supplementary materialThe online version of this article (10.1186/s40249-018-0439-y) contains supplementary material, which is available to authorized users.

show abstract

“…Some of the most frequently used have been kinetoplast DNA (kDNA) (15)(16)(17)(18), the GP63 gene (4), the β-tubulin gene (19), ITS rDNA (18,20), the miniexon gene (21)(22)(23)(24), and the gene that encodes heat shock protein 70 (hsp70) (2,25).…”

mentioning

confidence: 99%

Molecular typing of Leishmania (Leishmania) amazonensis and species of the subgenus Viannia associated with cutaneous and mucosal leishmaniasis in Colombia: A concordance study

et al. 2018

View full text Add to dashboard Cite

Introduction: Multilocus enzyme electrophoresis (MLEE) is the reference standard for the characterization of Leishmania species. The test is restricted to specialized laboratories due to its technical complexity, cost, and time required to obtain results. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is used to identify Leishmania species. Objective: To establish the concordance between the two tests as identifying methods for circulating species in Colombia. Materials and methods: A total of 96 isolates from patients with cutaneous or mucosal leishmaniasis were selected and identified by MLEE and PCR-RFLP with miniexon and hsp70 as the molecular targets, which were used sequentially. Restriction enzymes HaeIII and BccI were similarly applied. Cohen's kappa coefficient and the 95% confidence interval (CI) were calculated. Results: The kappa coefficient and the 95% CI between MLEE and PCR-RFLP displayed "very good" concordance with a coefficient of 0.98 (CI95%: 0.98 to 1.00). The identified species were Leishmania Viannia braziliensis, Leishmania Viannia panamensis, Leishmania Viannia guyanensis and Leishmania Leishmania amazonensis. A total of 80 of the 96 isolates were sequenced and the results obtained by PCR-RFLP were confirmed. Conclusion: Due to the concordance obtained between tests results with the amplification of the genes miniexon and hsp70, PCR-RFLP is proposed as an alternative for identifying circulating Leishmania species in Colombia.

show abstract

Intraspecific genetic variability in a population of Moroccan Leishmania infantum revealed by PCR-RFLP of kDNA minicircles

Cited by 13 publications

References 46 publications

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Genetic polymorphism in Leishmania infantum isolates from human and animals determined by nagt PCR-RFLP

Molecular typing of Leishmania (Leishmania) amazonensis and species of the subgenus Viannia associated with cutaneous and mucosal leishmaniasis in Colombia: A concordance study

Contact Info

Product

Resources

About