Copy number differences in the 195 bp repeated satellite DNA from Trypanosoma cruzi and Trypanosoma rangeli: potential use for epidemiologic surveys

Brenière, Simone Frédérique; Bosseno, M. F.; Barnabé, Christian; Morales, Servio Urdaneta; Tibayrenc, Michel

doi:10.1590/s0074-02761993000100027

Cited by 7 publications

(7 citation statements)

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“…Both OligoC-TesTs amplified purified T. rangeli DNA, but required between 10 2 and 10 7 times more template DNA than for T. cruzi . For the satDNA OligoC-TesT, this is probably due to the number of satDNA repeats being about 1000 times lower in T. rangeli than in T. cruzi [17]. For the kDNA OligoC-TesT, the higher detection limit for T. rangeli , was obtained by one base pair mismatch between the reverse primer and the kDNA sequence of T. rangeli .…”

Section: Discussionmentioning

confidence: 95%

The Trypanosoma cruzi Satellite DNA OligoC-TesT and Trypanosoma cruzi Kinetoplast DNA OligoC-TesT for Diagnosis of Chagas Disease: A Multi-cohort Comparative Evaluation Study

et al. 2014

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BackgroundThe Trypanosoma cruzi satellite DNA (satDNA) OligoC-TesT is a standardised PCR format for diagnosis of Chagas disease. The sensitivity of the test is lower for discrete typing unit (DTU) TcI than for TcII-VI and the test has not been evaluated in chronic Chagas disease patients.Methodology/Principal FindingsWe developed a new prototype of the OligoC-TesT based on kinetoplast DNA (kDNA) detection. We evaluated the satDNA and kDNA OligoC-TesTs in a multi-cohort study with 187 chronic Chagas patients and 88 healthy endemic controls recruited in Argentina, Chile and Spain and 26 diseased non-endemic controls from D.R. Congo and Sudan. All specimens were tested in duplicate. The overall specificity in the controls was 99.1% (95% CI 95.2%–99.8%) for the satDNA OligoC-TesT and 97.4% (95% CI 92.6%–99.1%) for the kDNA OligoC-TesT. The overall sensitivity in the patients was 67.9% (95% CI 60.9%–74.2%) for the satDNA OligoC-TesT and 79.1% (95% CI 72.8%–84.4%) for the kDNA OligoC-Test.Conclusions/SignificanceSpecificities of the two T. cruzi OligoC-TesT prototypes are high on non-endemic and endemic controls. Sensitivities are moderate but significantly (p = 0.0004) higher for the kDNA OligoC-TesT compared to the satDNA OligoC-TesT.

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Section: Discussionmentioning

confidence: 95%

The Trypanosoma cruzi Satellite DNA OligoC-TesT and Trypanosoma cruzi Kinetoplast DNA OligoC-TesT for Diagnosis of Chagas Disease: A Multi-cohort Comparative Evaluation Study

et al. 2014

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“…However, T. cruzi harbors satellite sequences at a much higher dosage than T. rangeli [38]. Moreover, leishmanial infections may lead to serological cross-reaction with T. cruzi .…”

Section: Discussionmentioning

confidence: 99%

Analytical Performance of a Multiplex Real-Time PCR Assay Using TaqMan Probes for Quantification of Trypanosoma cruzi Satellite DNA in Blood Samples

et al. 2013

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BackgroundThe analytical validation of sensitive, accurate and standardized Real-Time PCR methods for Trypanosoma cruzi quantification is crucial to provide a reliable laboratory tool for diagnosis of recent infections as well as for monitoring treatment efficacy.Methods/Principal FindingsWe have standardized and validated a multiplex Real-Time quantitative PCR assay (qPCR) based on TaqMan technology, aiming to quantify T. cruzi satellite DNA as well as an internal amplification control (IAC) in a single-tube reaction. IAC amplification allows rule out false negative PCR results due to inhibitory substances or loss of DNA during sample processing. The assay has a limit of detection (LOD) of 0.70 parasite equivalents/mL and a limit of quantification (LOQ) of 1.53 parasite equivalents/mL starting from non-boiled Guanidine EDTA blood spiked with T. cruzi CL-Brener stock. The method was evaluated with blood samples collected from Chagas disease patients experiencing different clinical stages and epidemiological scenarios: 1- Sixteen Venezuelan patients from an outbreak of oral transmission, 2- Sixty three Bolivian patients suffering chronic Chagas disease, 3- Thirty four Argentinean cases with chronic Chagas disease, 4- Twenty seven newborns to seropositive mothers, 5- A seronegative receptor who got infected after transplantation with a cadaveric kidney explanted from an infected subject.Conclusions/SignificanceThe performing parameters of this assay encourage its application to early assessment of T. cruzi infection in cases in which serological methods are not informative, such as recent infections by oral contamination or congenital transmission or after transplantation with organs from seropositive donors, as well as for monitoring Chagas disease patients under etiological treatment.

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“…14 The two parasites were distinguishable by their morphology and biological behavior but few specific genetic markers have been proposed. 15,16,17,18 Moreover, mammals can harbor several species of the subgenus Schizotrypanum and genetic characterization should help to determine their importance as a reservoir for T. cruzi.…”

Section: Introductionmentioning

confidence: 99%

Isoenzyme clustering of Trypanosomatidae Colombian populations.

Montilla¹,

GUHL²,

Jaramillo³

et al. 2002

The American Journal of Tropical Medicine and Hygiene

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Abstract. Thirty-six Trypanosomatidae stocks isolated from various hosts and geographical areas in Colombia and 7 others from Bolivia, Chile, Honduras and Panama have been surveyed by Multilocus Enzyme Electrophoresis (MLEE). Part of the Colombian stocks were previously characterized by morphology and biological behavior as belonging to Trypanosoma cruzi and Trypanosoma rangeli taxa, others were unknown species. The genetic variability observed at 13 different loci was considerable, since 38 zymodemes could be distinguished and 2 upper branches were observed. The first branch corresponded to T. cruzi and was divided in the two major phylogenetic subdivision of T. cruzi (T. cruzi I, T. cruzi II). The majority of the Colombian T. cruzi stocks (92%) felt into T. cruzi I. Only two stocks, isolated from sylvatic mammals, belonged to T. cruzi II. Among T. cruzi I, we did not observed any additional phylogenetic subdivision and host-dependent genotype specificity. The second branch was genetically very heterogeneous and included all T. rangeli stocks, the stocks isolated from bats and one stock isolated from a sylvatic R. prolixus vector. The stocks belonging to T. rangeli presented only one locus instead of two for the malic enzyme system. Since, the upper level of resolution of the isoenzyme method was exceeded, the current clustering study failed to draw a clear distinction between such a diverse set of Trypanosomatidae species.

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Copy number differences in the 195 bp repeated satellite DNA from Trypanosoma cruzi and Trypanosoma rangeli: potential use for epidemiologic surveys

Cited by 7 publications

References 0 publications

The Trypanosoma cruzi Satellite DNA OligoC-TesT and Trypanosoma cruzi Kinetoplast DNA OligoC-TesT for Diagnosis of Chagas Disease: A Multi-cohort Comparative Evaluation Study

The Trypanosoma cruzi Satellite DNA OligoC-TesT and Trypanosoma cruzi Kinetoplast DNA OligoC-TesT for Diagnosis of Chagas Disease: A Multi-cohort Comparative Evaluation Study

Analytical Performance of a Multiplex Real-Time PCR Assay Using TaqMan Probes for Quantification of Trypanosoma cruzi Satellite DNA in Blood Samples

Isoenzyme clustering of Trypanosomatidae Colombian populations.

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