2.3.3. Characterisation of the chronic CD
15 infected mice were used to characterise the chronic CD in a
time-dependent manner, sacrificing 5 mice by cervical dislocation for
each experimental time: 20th, 30th,
and 60th dpi. 14 organs/tissues were harvested to
evaluate the tropism: adipose tissue, bone marrow, brain, oesophagus,
heart, kidney, large intestine, liver, lung, mesenteric tissue, muscle,
small intestine, spleen and stomach (Scheme 1 ). Hearts and
spleens were also weighed to evaluate splenomegaly and cardiomegaly.
These organs/tissues were immediately flushed free of blood by infusion
of pre-warmed PBS to avoid contamination with BTs, and they were then
thawed and ground up using a Potter-Elvehjem. Organs/tissues DNA was
extracted using Wizard Genomic DNA Purification Kit (Promega)
(MartÃn-Escolano et al. , 2018). PCR was based on the Spliced
Leader (SL) intergenic region sequence of T. cruzi (for detailed
description, see Paucar et al. (Paucar et al. , 2019). Finally,
the PCR products were visualised by UV illumination after a 2 % agarose
gel electrophoresis for 90 min at 90 V, containing 1:10000 GelRed
nucleic acid gel stain.