Towards a standard battery of microsatellite markers for the analysis of the<i>Leishmania donovani</i>complex

Jamjoom, Manal B.; Ashford, R. W.; Bates, Paul A.; Kemp, Stephen J.; Noyes, H. A.

doi:10.1179/000349802125000790

Cited by 52 publications

(43 citation statements)

References 13 publications

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“…The 20 microsatellite loci investigated (15 already published [26] and five developed in the laboratory) are listed in Supplementary data Table S1. The 61 strains (and M9702, as L. chagasi outgroup) under study were amplified according to the following conditions.…”

Section: Methodsmentioning

confidence: 99%

Multifaceted Population Structure and Reproductive Strategy in Leishmania donovani Complex in One Sudanese Village

et al. 2011

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Leishmania species of the subgenus Leishmania and especially L. donovani are responsible for a large proportion of visceral leishmaniasis cases. The debate on the mode of reproduction and population structure of Leishmania parasites remains opened. It has been suggested that Leishmania parasites could alternate different modes of reproduction, more particularly clonality and frequent recombinations either between related individuals (endogamy) or between unrelated individuals (outcrossing) within strongly isolated subpopulations. To determine whether this assumption is generalized to other species, a population genetics analysis within Leishmania donovani complex strains was conducted within a single village. The results suggest that a mixed-mating reproduction system exists, an important heterogeneity of subsamples and the coexistence of several genetic entities in Sudanese L. donovani. Indeed, results showed significant genetic differentiation between the three taxa (L. donovani, L. infantum and L. archibaldi) and between the human or canine strains of such taxa, suggesting that there may be different imbricated transmission cycles involving either dogs or humans. Results also are in agreement with an almost strict specificity of L. donovani stricto sensu to human hosts. This empirical study demonstrates the complexity of population structure in the genus Leishmania and the need to pursue such kind of analyses at the smallest possible spatio-temporal and ecological scales.

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Section: Methodsmentioning

confidence: 99%

Multifaceted Population Structure and Reproductive Strategy in Leishmania donovani Complex in One Sudanese Village

et al. 2011

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“…It will also help to answer specific clinical questions, such as the role of parasite persistence after subclinical infection; whether endogenous versus exogenous reinfection is associated with immunosuppression; and whether transmission is vector independent, such as via blood transfusion or organ transplantation. In Leishmania species, microsatellite markers are specific at the species or species complex level (16,31). When DNA from other species of Leishmania is used, in most cases no PCR product is obtained or the products differ significantly in size.…”

Section: Discussionmentioning

confidence: 99%

Development of a Multilocus Microsatellite Typing Approach for Discriminating Strains of Leishmania ( Viannia ) Species

et al. 2009

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A multilocus microsatellite typing (MLMT) approach based on the analysis of 15 independent loci has been developed for the discrimination of strains belonging to different Viannia species. Thirteen microsatellite loci were isolated de novo from microsatellite-enriched libraries for both Leishmania braziliensis and L. guyanensis. Two previously identified markers, AC01 and AC16, were modified and added to our marker set. Cutaneous leishmaniasis (CL) is a serious but neglected public health problem that is endemic in 22 countries of Latin America, with Brazil and Colombia being among the six countries reporting 90% of the worldwide cases. Species of the subgenus Viannia cause the majority of cases of CL and mucocutaneous leishmaniasis (MCL) in South America and often overlap in their distribution (22). The subgenus is subdivided into species complexes representing Leishmania braziliensis, L. guyanensis, and L. naiffi (8). The L. guyanensis complex includes some named species, such as L. panamensis and L. shawi, which were found to be very similar to each other (7).

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“…A high number of independent microsatellite markers developed by us and other authors (5,22,47) are now available for multilocus microsatellite typing (MLMT) (12) of the L. donovani complex. In order to make high-throughput typing feasible, optimization of fragment analysis is most important.…”

Section: Discussionmentioning

confidence: 99%

Multilocus Microsatellite Typing as a New Tool for Discrimination of Leishmania infantum MON-1 Strains

et al. 2006

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The Leishmania donovani complex, which consists of L. donovani, L. infantum-L. chagasi, and L. archibaldi, is responsible for visceral manifestations of leishmaniasis. Multilocus enzyme electrophoresis is the standard method for the characterization and identification of strains of Leishmania. For L. infantum, the predominance of zymodeme MON-1 significantly reduces the discriminative power of this approach. In the present study, we developed 17 independent polymorphic microsatellite markers for the typing of strains of L. infantum, with the main emphasis on zymodeme MON-1. The discriminative powers of 11 markers selected from among these markers were tested by using a panel of 63 isolates of the L. donovani complex. Unique multilocus genotypes were observed for the strains analyzed, with only three exceptions. Model-based and distance-based analyses of the data set showed comparable results. It was possible to discriminate between L. donovani sensu stricto, a non-MON-1 group of L. infantum isolates, and a MON-1 group of L. infantum isolates. Within MON-1, three clusters with geographical correlations became apparent. The frequency of heterozygosity in the alleles analyzed varied extremely between the different groups of isolates. The main clusters described are not consistent with species definitions based on isoenzyme analysis but confirm the results of former PCR-based investigations.Leishmania is a genus of protozoan flagellates that cause a broad spectrum of diseases, ranging from self-limiting localized cutaneous lesions to visceral leishmaniasis with fatal spontaneous evolution (2). The majority of visceral manifestations are caused by parasites of the Leishmania donovani complex (26), which consists of L. donovani Ross, 1903; L. infantum Nicolle, 1908-L. chagasi Cunha and Chagas, 1937; and L. archibaldi Castellani and Chalmers, 1919. Currently, multilocus enzyme electrophoresis (MLEE) is the generally accepted "gold standard" for the identification and classification of isolates of Leishmania. By this method, strains are divided into groups with identical enzyme patterns, called "zymodemes." The main criticism of this approach is that genotypes are assayed indirectly, with the consequence that nucleotide substitutions may not be observed in synonymous sites or in nonsynonymous sites, if it is assumed that subsequent changes in the amino acid composition do not lead to different electrophoretic mobilities. In contrast, posttranslational modifications may change the electrophoretic mobilities, despite identical genotypes. Furthermore, the method is quite slow, laborious, and costly. Growth in vitro is inevitable, and the data sets of the few laboratories in which these analyses are performed are difficult to compare.Unlike, e.g., L. donovani or L. tropica, which present extended genetic and enzymatic polymorphisms (27, 53), L. infantum is a relatively uniform species (46). More than 80% of the strains of L. infantum isolated thus far belong to the predominant zymodeme, 42). The alternative methods for ...

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Towards a standard battery of microsatellite markers for the analysis of theLeishmania donovanicomplex

Cited by 52 publications

References 13 publications

Multifaceted Population Structure and Reproductive Strategy in Leishmania donovani Complex in One Sudanese Village

Multifaceted Population Structure and Reproductive Strategy in Leishmania donovani Complex in One Sudanese Village

Development of a Multilocus Microsatellite Typing Approach for Discriminating Strains of Leishmania ( Viannia ) Species

Multilocus Microsatellite Typing as a New Tool for Discrimination of Leishmania infantum MON-1 Strains

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