2.5. Detection of Leishmania tropica DNA
The presence of L. tropica DNA was investigated in P.
sergenti females captured in both localities, El Borouj and Sidi
Hajjaj, using Granaleish Multiplex qPCR (University of Granada, Spain,
Trade Mark Number 3667362/5). This PCR technique can differentiate
between L. infantum, L. tropica and L. major and allows
quantification of the parasite load (Merino-Espinosa et al.,2018). Primers F, R and the 3 Taqman probes were provided by the
manufacturer. The following thermal profile was used: 10 min at 95 °C,
then 36 cycles of 30 s at 95 °C and 60 s at 60 °C. The number of
parasites in every qPCR reaction was calculated through the
interpolation of the cycle threshold (Ct) value in a standard curve.