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. 2010 Jan;82(1):67-73.
doi: 10.4269/ajtmh.2010.09-0391.

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)

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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)

Cristina Diez et al. Am J Trop Med Hyg. 2010 Jan.

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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Figures

Figure 1.
Figure 1.
Agarose gel of minicircle lineage-specific polymerase chain reaction (MLS-PCR) performed with some reference strains of all Trypanosoma cruzi sublineages. 1.5% agarose gel Etidium Bromide-stained. Lanes: 1) Negative Control, 2) X10 cl1 (TcI), 3) CANIII cl1 (TcIIa), 4) CBB (TcIIb), 5) IVV cl4 (TcIIb), 6) M5631 cl5 (TcIIc), 7) MNcl2 (TcIId), 8) XHCH56 (TcIId), 9) NR cl3(TcIId), 10) JGG (TcIId), 11) SC43cl1(TcIId), 12) V195 cl1(TcIIe), 13) CL Brener (TcIIe).
Figure 2.
Figure 2.
Infections with different Trypanosoma cruzi genotypes analyzed in the congenital and chronic samples performed with hybridization probes. Black bar indicates chronic samples and white bar indicates congenital samples. TcIId and TcIIe correspond to single infections with these sublinage, Mixed TcIId/e or TcIIb/e correspond to combined infections and Not Genotyped correspond to samples of patients that did not hybridize with any probe tested.

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