In visceral leishmaniasis, the detection of the agent is of paramount importance to
identify reservoirs of infection. Here, we evaluated the diagnostic attributes of
PCRs based on primers directed to cytochrome-B (cytB),
cytochrome-oxidase-subunit II (coxII), cytochrome-C
(cytC), and the minicircle-kDNA. Although PCRs directed to
cytB, coxII, cytC were able to detect different species of
Leishmania, and the nucleotide sequence of their amplicons
allowed the unequivocal differentiation of species, the analytical and diagnostic
sensitivity of these PCRs were much lower than the analytical and diagnostic
sensitivity of the kDNA-PCR. Among the 73 seropositive animals, the asymptomatic dogs
had spleen and bone marrow samples collected and tested; only two animals were
positive by PCRs based on cytB, coxII, and
cytC, whereas 18 were positive by the kDNA-PCR. Considering the
kDNA-PCR results, six dogs had positive spleen and bone marrow samples, eight dogs
had positive bone marrow results but negative results in spleen samples and, in four
dogs, the reverse situation occurred. We concluded that PCRs based on
cytB, coxII, and cytC can be
useful tools to identify Leishmania species when used in combination
with automated sequencing. The discordance between the results of the kDNA-PCR in
bone marrow and spleen samples may indicate that conventional PCR lacks sensitivity
for the detection of infected dogs. Thus, primers based on the kDNA should be
preferred for the screening of infected dogs.
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