Trypanosoma cruzi: Sequence analysis of the variable region of kinetoplast minicircles

Tellería, Jenny; Lafay, Bénédicte; Virreira, Myrna; Barnabé, Christian; Tibayrenc, Michel; Svoboda, Michal

doi:10.1016/j.exppara.2006.04.005

Cited by 67 publications

(48 citation statements)

References 47 publications

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“…According to some authors (17,34,36), the genetic variability of strains of T. cruzi I can lead to the inference that a difference may exist between the kDNA sequences of strains belonging to the T. cruzi I lineage and those belonging to the T. cruzi II lineage. Phylogenetic analysis based on the variable region of T. cruzi kDNA demonstrated genetic variability when isolates belonging to the T. cruzi I and II lineages were compared (37). Also, the influence of the stDNA copy number difference between the T. cruzi lineages has been established (12,15,24), where the number of stDNA copies in T. cruzi I is lower than the number in T. cruzi II.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Adult Chronic Chagas' Heart Disease Diagnosis by Molecular and Serological Methods

et al. 2009

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Chagas' disease caused by Trypanosoma cruzi is endemic in Latin America. T. cruzi presents heterogeneous populations and comprises two main genetic lineages, named T. cruzi I and T. cruzi II. Diagnosis in the chronic phase is based on conventional serological tests, including indirect immunofluorescence (IIF) and enzymelinked immunosorbent assay (ELISA), and diagnosis in the acute phase based on parasitological methods, including hemoculture. The objective of this study was to evaluate the diagnostic procedures of Chagas' disease in adult patients in the chronic phase by using a PCR assay and conventional serological tests, including TESA-blot as the gold standard. Samples were obtained from 240 clinical chronic chagasic patients. The sensitivities, compared to that of TESA-blot, were 70% for PCR using the kinetoplast region, 75% for PCR using the nuclear repetitive region, 99% for IIF, and 95% for ELISA. According to the serological tests results, we recommend that researchers assess the reliability and sensitivity of the commercial kit Chagatest ELISA recombinant, version 3.0 (Chagatest Rec v3.0; Wiener Lab, Rosario, Argentina), due to the lack of sensitivity. Based on our analysis, we concluded that PCR cannot be validated as a conventional diagnostic technique for Chagas' disease. These data have been corroborated by low levels of concordance with serology test results. It is recommended that PCR be used only for alternative diagnostic support. Using the nuclear repetitive region of T. cruzi, PCR could also be applicable for monitoring patients receiving etiologic treatment.

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Section: Discussionmentioning

confidence: 99%

Evaluation of Adult Chronic Chagas' Heart Disease Diagnosis by Molecular and Serological Methods

et al. 2009

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“…24 On the other hand, two pairs of primers amplify predominant sequences in TcIId sublineage, covering 80% of sequences found in that sublineage. 23 In a previous work, using specific probes for two TcIId predominant sequences (O and P), Virreira and others proposed a subclassification within this sublineage. In this way, TcIId were subdivided in MN-like, Bug2148-like, and TPK-like, if amplified PCR products hybridized mainly with O sequence, mainly with P, or equally with both of them, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…8,[17][18][19][20][21][22] Beyond the high variability of mHVR, the comparison of several minicircle regions within and among strains, has allowed identification of some sequences more represented in certain sublineages. 23 In a previous work, 24 we have described that a predominant minicircle sequence present in TcIIe and two others predominant minicircle sequences present in TcIId, are indeed present in reference strains from other T. cruzi lineages and sublineages . However, the amount of these sequences is at least a thousand times more represented in the respective sublineage than in the others.…”

Section: Introductionmentioning

confidence: 99%

“…However, the amount of these sequences is at least a thousand times more represented in the respective sublineage than in the others. Supported by this fact, we have developed a semiquantitative PCR directly from biological samples, using GrpI primers, designed by Velázquez and others, 24 and primers (GrO-GrP) that amplify O and P sequences, 23 to discriminate TcIIe and TcIId sublineages, respectively, and to differentiate them from the other lineages and sublineages. In this work, we have evaluated this assay by extending the number of reference strains analyzed and testing biological samples by hybridization with a panel of genotypespecific minicircle DNA probes.…”

Section: Introductionmentioning

confidence: 99%

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Genotyping of Trypanosoma cruzi Sublineage in Human Samples from a North-East Argentina Area by Hybridization with DNA Probes and Specific Polymerase Chain Reaction (PCR)

Diez¹,

Lorenz²,

Ortíz³

et al. 2010

The American Journal of Tropical Medicine and Hygiene

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Abstract. We have evaluated blood samples of chronic and congenital Trypanosoma cruzi -infected patients from the city of Reconquista located in the northeast of Argentina where no information was previously obtained about the genotype of infecting parasites. Fourteen samples of congenital and 19 chronical patients were analyzed by hybridization with DNA probes of minicircle hypervariable regions (mHVR). In congenital patients, 50% had single infections with TcIId, 7% single infections with TcIIe, 29% mixed infections with TcIId/e, and 7% had mixed infections with TcIId/b and 7% TcIId/b, respectively. In Chronical patients, 52% had single infections with TcIId, 11% single infections with TcIIe, 26% had mixed infections with TcIId/e, and 11% had non-identified genotypes. With these samples, we evaluated the minicircle lineage-specific polymerase chain reaction assay (MLS-PCR), which involves a nested PCR to HVR minicircle sequences and we found a correlation with hybridization probes of 96.4% for TcIId and 54.8% for TcIIe.

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“…These lineages appear to be distributed differentially among triatomine bugs, vertebrate host species and habitats in different geographical areas (Higo et al, 2004, Yeo et al, 2005. Although all T. cruzi lineages cause human disease, some studies suggest that T. cruzi IIb, IId and IIe are more likely to be associated with anthropic environments and chronic Chagas disease patients, T. cruzi IIa and IIc to sylvatic environments, and T. cruzi I to both (Telleria et al, 2006;Yeo et al, 2005). Although there is a wealth of studies on the genetic diversity of T. cruzi populations, there has been a tendency to re-examine the available parasite isolates.…”

Section: Introductionmentioning

confidence: 99%

Molecular epidemiology of domestic and sylvatic Trypanosoma cruzi infection in rural northwestern Argentina

Cardinal

Lauricella

Ceballos

et al. 2008

International Journal for Parasitology

106

104

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Genetic diversity of Trypanosoma cruzi populations and parasite transmission dynamics have been well documented throughout the Americas, but few studies have been conducted in the Gran Chaco ecoregion, one of the most highly endemic areas for Chagas disease, caused by T. cruzi. In this study we assessed the distribution of T. cruzi lineages (identified by PCR strategies) in Triatoma infestans, domestic dogs, cats, humans and sylvatic mammals from two neighboring rural areas with different histories of transmission and vector control in northern Argentina. Lineage II predominated among the 99 isolates characterized and lineage I among the six isolates obtained from sylvatic mammals. Trypanosoma cruzi lineage IIe predominated in domestic habitats; it was found in 87% of 54 isolates from Tr. infestans, in 82% of 33 isolates from dogs, and in the four cats found infected. Domestic and sylvatic cycles overlapped in the study area in the late 1980s, when intense domestic transmission occurred, and still overlap marginally. The introduction of T. cruzi from sylvatic into domestic habitats is likely to occur very rarely in the current epidemiological context. The household distribution of T. cruzi lineages showed that Tr. infestans, dogs and cats from a given house compound shared the same parasite lineage in most cases. Based on molecular evidence, this result lends further support to the importance of dogs and cats as domestic reservoir hosts of T. cruzi. We believe that in Argentina, this is the first time that lineage IIc has been isolated from naturally-infected domestic dogs and Tr. infestans.

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Trypanosoma cruzi: Sequence analysis of the variable region of kinetoplast minicircles

Cited by 67 publications

References 47 publications

Evaluation of Adult Chronic Chagas' Heart Disease Diagnosis by Molecular and Serological Methods

Evaluation of Adult Chronic Chagas' Heart Disease Diagnosis by Molecular and Serological Methods

Genotyping of Trypanosoma cruzi Sublineage in Human Samples from a North-East Argentina Area by Hybridization with DNA Probes and Specific Polymerase Chain Reaction (PCR)

Molecular epidemiology of domestic and sylvatic Trypanosoma cruzi infection in rural northwestern Argentina

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