RAPD in the analysis of isolates of Entamoeba histolytica

Gomes, Maria Aparecida; Melo, Maria Norma; Macedo, Andréa Mara; Furst, Cinthia; Silva, Edward F.

doi:10.1016/s0001-706x(99)00097-2

Cited by 12 publications

(6 citation statements)

References 21 publications

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“…Hadrys et al (1992) considered that the applications for RAPD included species identi®cation. RAPD markers analyzed with clustering methods, such as UPGMA, permitted isolates or individuals to be grouped according to species (Tibayrenc et al 1993;Gomes et al 2000). Our data con®rm the usefulness of these techniques in distinguishing between Eimeria isolates, according to species.…”

Section: Discussionsupporting

confidence: 60%

“…Gomes et al (1995), studying Leishmania (Viannia) braziliensis in Brazil, observed that isolates from the State of Minas Gerais formed a cluster which was genetically isolated from those originating from the State of ParaÂ . Using the same approach to study isolates of Entamoeba spp, Gomes et al (2000) built a phenogram in which the isolates formed three clusters: one with the E. invadens isolate, another with two E. moshkovskii isolates, and a third cluster with the 11 E. histolytica isolates.…”

Section: Introductionmentioning

confidence: 99%

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Eimeria parasites of domestic fowl: genetic relationships of different isolates estimated from random amplified polymorphic DNA

Costa

Gomes

Melo

et al. 2001

Parasitology Research

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Genetic relationships between Eimeria parasites of domestic fowl, including three isolates of E. acervulina, one of E. maxima, four of E. mitis, six of E. praecox, one of E. tenella, and two of uncertain identity, were analyzed by random amplified polymorphic DNA (RAPD) and the unweighted pair-group method with arithmetic mean, using 12 primers. Each primer amplified 4-34 DNA fragments/isolate. The similarity coefficients and phenograms were calculated from RAPD products with 230-2,000 bp in silver-stained polyacrylamide gels. Some primers generated RAPD markers which were species-specific for E. acervulina, E. mitis, and E. praecox. The phenograms revealed six clusters, each corresponding to an individual Eimeria species. The pBP and pBP2 isolates, of uncertain identity, clustered with the E. acervulina isolates (100% bootstrap). The intraspecific relationships showed certain degree of genetic isolation of the Eimeria populations, and it was associated with broiler house and with geographic separation.

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Section: Discussionsupporting

confidence: 60%

Section: Introductionmentioning

confidence: 99%

Eimeria parasites of domestic fowl: genetic relationships of different isolates estimated from random amplified polymorphic DNA

Costa

Gomes

Melo

et al. 2001

Parasitology Research

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“…Gomes et al (2000) have reported RAPD analysis using axenic strains of E. histolytica. Using 11 reference strains of E. histolytica originally isolated from 4 asymptomatic and 7 symptomatic individuals, mainly from Brazil and Mexico, the authors found that only about 70% of the RAPD bands were shared.…”

Section: Polymorphism In Non-coding Dnamentioning

confidence: 99%

Molecular epidemiology of amebiasis

Ali

Clark

Petri

2008

Infection, Genetics and Evolution

127

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Entamoeba histolytica, the causative agent of human amebiasis, remains a significant cause of morbidity and mortality in developing countries and is responsible for up to 100,000 deaths worldwide each year. Entamoeba dispar, morphologically indistinguishable from E. histolytica, is more common in humans in many parts of the world. Similarly Entamoeba moshkovskii, which was long considered to be a free-living ameba, is also morphologically identical to E. histolytica and E. dispar, and is highly prevalent in some E. histolytica endemic countries. However, the only species to cause disease in humans is E. histolytica. Most old epidemiological data on E. histolytica are unusable as the techniques employed do not differentiate between the above three Entamoeba species. Molecular tools are now available not only to diagnose these species accurately but also to study intra-species genetic diversity. Recent studies suggest that only a minority of all E. histolytica infections progress to development of clinical symptoms in the host and there exist population level differences between the E. histolytica strains isolated from the asymptomatic and symptomatic individuals. Nevertheless the underlying factors responsible for variable clinical outcome of infection by E. histolytica remain largely unknown. We anticipate that the recently completed E. histolytica genome sequence and new molecular techniques will rapidly advance our understanding of the epidemiology and pathogenicity of amebiasis.

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“…Studies have analyzed a small number of highly repetitive and polymorphic genetic loci by techniques, such as randomly amplified polymorphic DNA (RAPD), RNA arbitrarily primed PCR, and restriction fragment length polymorphism (RFLP), to conclude that there is significant genetic diversity (7,18,23,31,33,64,72,73). Contradictory data were obtained by two studies; in one study, the lectin gene was sequenced, and in the other study, the intergenic region between the superoxide dismutase gene and the actin 3 gene was analyzed (9,30).…”

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confidence: 99%

Comparative Genomic Hybridizations ofEntamoebaStrains Reveal Unique Genetic Fingerprints That Correlate with Virulence

et al. 2005

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Variable phenotypes have been identified for Entamoeba species. Entamoeba histolytica is invasive and causes colitis and liver abscesses but only in ϳ10% of infected individuals; 90% remain asymptomatically colonized. Entamoeba dispar, a closely related species, is avirulent. To determine the extent of genetic diversity among Entamoeba isolates and potential genotype-phenotype correlations, we have developed an E. histolytica genomic DNA microarray and used it to genotype strains of E. histolytica and E. dispar. On the basis of the identification of divergent genetic loci, all strains had unique genetic fingerprints. Comparison of divergent genetic regions allowed us to distinguish between E. histolytica and E. dispar, identify novel genetic regions usable for strain and species typing, and identify a number of genes restricted to virulent strains. Among the four E. histolytica strains, a strain with attenuated virulence was the most divergent and phylogenetically distinct strain, raising the intriguing possibility that genetic subtypes of E. histolytica may be partially responsible for the observed variability in clinical outcomes. This microarray-based genotyping assay can readily be applied to the study of E. histolytica clinical isolates to determine genetic diversity and potential genotypic-phenotypic associations.Entamoeba histolytica causes amebic colitis and liver abscesses. Worldwide, 50 million people have invasive E. histolytica, and 100,000 die each year, making it the second most common cause of parasitic death in humans (71). In Dhaka, Bangladesh, where diarrheal diseases are the leading cause of death in children less than 6 years old, ϳ50% of children have serological evidence of exposure to E. histolytica by age 5 (34, 35). Infected children suffer from significant morbidity with malnourishment and growth delays. A number of interesting epidemiologic trends have been identified for amebic disease. The data, although limited, indicate that a minority (ϳ10%) of individuals who become infected with E. histolytica progress to clinically overt disease; others remain asymptomatically colonized (29). In individuals with invasive disease, occurrence of amebic liver abscess (ALA) is 5 to 50 times less common than diarrhea (65). Geographically variable disease predilections have been observed, and invasive disease predominantly affects men (1, 2, 11, 61).The extent of genetic diversity among E. histolytica clinical isolates is unclear. Studies have analyzed a small number of highly repetitive and polymorphic genetic loci by techniques, such as randomly amplified polymorphic DNA (RAPD), RNA arbitrarily primed PCR, and restriction fragment length polymorphism (RFLP), to conclude that there is significant genetic diversity (7,18,23,31,33,64,72,73). Contradictory data were obtained by two studies; in one study, the lectin gene was sequenced, and in the other study, the intergenic region between the superoxide dismutase gene and the actin 3 gene was analyzed (9, 30). These studies found minimal genetic diversi...

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RAPD in the analysis of isolates of Entamoeba histolytica

Cited by 12 publications

References 21 publications

Eimeria parasites of domestic fowl: genetic relationships of different isolates estimated from random amplified polymorphic DNA

Eimeria parasites of domestic fowl: genetic relationships of different isolates estimated from random amplified polymorphic DNA

Molecular epidemiology of amebiasis

Comparative Genomic Hybridizations ofEntamoebaStrains Reveal Unique Genetic Fingerprints That Correlate with Virulence

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