Journal: Microbiology Spectrum

Article Title: TIM, a novel molecular toolbox for the detection and identification of Leishmania species in Morocco

doi: 10.1128/spectrum.03447-25

Figure Lengend Snippet: ( A ) Schematic representation of nested PCR on kDNA minicircles developed by Noyes et al. and ITS1-RFLP on rDNA developed by Schönian et al. . Arrows represent primers’ location and names. Sizes of the expected amplification products (Noyes) or Hae III restriction fragments (ITS1-RFLP) are shown on the right. ( B ) Overview of the three TIM1, TIM2, and TIM3 tests with a schematic representation of a minicircle and its conserved domain (rectangle). Primers location and names are indicated with arrows. Virtual migration of amplified products is shown at the bottom. Ltrop, Lt: L. tropica ; Linf, Li : L. infantum ; Lmaj, Lm: L. major . ( C ) Sizes, names, and relative positions of primers used on both Noyes and TIM nested tests from the conserved domain of minicircles. CSB: conserved sequence block 1–3.

Article Snippet: Next, 15 μL of PCR products was digested with the Hae III (New England Biolabs) at 37°C for 1 h. The species were identified according to the restriction fragment sizes observed : L. major (two bands at 220 and 127 bp), L. tropica (two bands at 220 and 50 bp), and L. infantum (three bands at 200, 100, and 50 bp).

Techniques: Nested PCR, Amplification, Migration, Sequencing, Blocking Assay