Figure 1.
Dynamic range of the T. cruzi satellite DNA based Q-PCR.
Results are expressed as the number of parasites per milliliter of blood and represent the average of 5 independent experiments. Slope = −3.35. Efficiency = 99%. Dynamic range: 0.01–105 p/mL. R square: 0.998. C(t): cycle threshold.
Table 1.
Variation in the relative numbers of satellite DNA copies per genome.
Figure 2.
Melting curve analysis of satellite amplicons from reference stocks.
Group I satellite amplicons show melting temperatures above 85°C, whereas Group II render amplification products with melting temperatures below 85°C.
Table 2.
Melting temperatures of satellite fragments amplified from T. cruzi stocks belonging to the 6 phylogenetic lineages.
Table 3.
Examples of the calculation of parasitic loads in pediatric and transplanted patients.
Table 4.
Reproducibility of the Q-PCR and IS-PCR assays in clinical samples of Chagas disease patients.
Figure 3.
Parasitic loads in peripheral blood samples from pediatric patients.
(A) Association between basal parasitic loads and patients' ages in 43 pediatric cases. Coefficient of correlation: −0.5832; P<0.05. (B) Monitoring of parasitological response to benznidazole therapy in 38 pediatric patients. The evolution of the parasitic loads for patients with more then one positive sample are depicted. Samples were withdrawn at time of diagnosis (t1), after 7 (t2) and 30 (t3) days of treatment, as well as at the end of treatment (t4, 60 days). Only the PCR positive samples are shown. The horizontal line represents the lower limit of the dynamic range of Q-PCR.
Figure 4.
Follow-up of Chronic Chagas heart disease patients after heart transplantation.
Parasitic loads in peripheral blood samples of Chronic Chagas heart disease patients with clinical reactivation due to immunosupression after heart transplantation. * Time of diagnosis of clinical reactivation and etiological treatment.