Genetic profiling of Trypanosoma cruzi directly in infected tissues using nested PCR of polymorphic microsatellites

Trypanosoma cruzi: parasite persistence in tissues in chronic chagasic Brazilian patients

“…Our data emphasizes that molecular biology tools, such as NPCR and real-time PCR, used to detect and to quantify DNA extracted from fixed tissues may contribute to a better comprehension of the pathogenesis of Chagas disease (Valadares et al 2008). The difference in the number of copies found in different tissues by realtime PCR indicates that the variability in parasite load is associated with pathological alterations in target organs as well with disease progression.…”

Trypanosoma cruzi benznidazole susceptibility in vitro does not predict the therapeutic outcome of human Chagas disease

“…Total parasite DNA was obtained as described (Macedo et al 1992). T. cruzi genotyping was performed by amplifying nine microsatellite loci: SCLE10, SCLE11, MCLE01, MCLF10 and MCLG10 (Oliveira et al 1998) and TcTAT20, TcAAT8, TcATT14 and TcAAAT6 (Valadares et al 2007). Amplification of microsatellites by PCR was achieved as previously described (Oliveira et al 1998, Valadares et al 2007).…”

“…T. cruzi genotyping was performed by amplifying nine microsatellite loci: SCLE10, SCLE11, MCLE01, MCLF10 and MCLG10 (Oliveira et al 1998) and TcTAT20, TcAAT8, TcATT14 and TcAAAT6 (Valadares et al 2007). Amplification of microsatellites by PCR was achieved as previously described (Oliveira et al 1998, Valadares et al 2007). The microsatellite allele sizes were determined using the Allelelocator software (GE Healthcare) after running 1-3 µL of PCR fluorescent amplicons in a 6% denaturing polyacrylamide gel on an ALF sequencer (GE Healthcare, Milwaukee, Wisconsin, USA) in comparison to standard size DNA fragments of 50-500 bp.…”

The molecular epidemiology and phylogeography of Trypanosoma cruzi and parallel research on Leishmania: looking back and to the future

“…Ideally, the conundrum of whether infecting genotypes, as well as host factors, govern clinical manifestations (Carranza et al 2009) can be addressed, although this hope is yet to be fully realised and may require innovative technical approaches to reveal the T. cruzi genotypes present in blood and internal organs (Vago et al 2000 ;Valadares et al 2008) and to resolve the patient's prior history of exposure to such genotypes.…”