Progress in the diagnosis of leishmaniases depends on the development of effective methods and the discovery of suitable biomarkers. We propose firstly an update classification of Leishmania species and their synonymies. We demonstrate a global map highlighting the geography of known endemic Leishmania species pathogenic to humans. We summarize a complete list of techniques currently in use and discuss their advantages and limitations. The available data highlights the benefits of molecular markers in terms of their sensitivity and specificity to quantify variation from the subgeneric level to species complexes, (sub) species within complexes, and individual populations and infection foci. Each DNA-based detection method is supplied with a comprehensive description of markers and primers and proposal for a classification based on the role of each target and primer in the detection, identification and quantification of leishmaniasis infection. We outline a genome-wide map of genes informative for diagnosis that have been used for Leishmania genotyping. Furthermore, we propose a classification method based on the suitability of well-studied molecular markers for typing the 21 known Leishmania species pathogenic to humans. This can be applied to newly discovered species and to hybrid strains originating from inter-species crosses. Developing more effective and sensitive diagnostic methods and biomarkers is vital for enhancing Leishmania infection control programs.

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During 2013–2015, prevalence of cutaneous leishmaniasis in war-affected Waziristan areas was 3.61% by PCR. Youths (1–15 years of age) were more susceptible. Internal transcribed spacer 1 PCR followed by restriction fragment length polymorphism analysis identified Leishmania tropica in 215 samples and Leishmania major in 6 samples.

Dogs are the major reservoir ofLeishmania infantum, the causative agent of human and canine visceral leishmaniasis in the Mediterranean basin. In Morocco, canine leishmaniasis (CanL) is usually believed to be widespread mainly, if not only, in the northern regions and few data are available about the situation in southern parts of the country. Here, we report the results of a preliminary, clinical, and serological study carried out in 2004–2007, in four provinces of southern Morocco. Serological analyses were processed using two different Elisa techniques, a homemade Elisa test and IDVET commercial kit, and confirmed by two different western blot (WB) tests, homemade and LDBIO commercial kits. We highlighted the presence of CanL infection in southern regions, known until then as free of the disease: 19.8% (48/243) of examined dogs displayed clinical signs compatible with CanL and the seroprevalence was particularly high, respectively, 81.8% and 87.8% by Elisa and western blot tests. Our current developed and validated homemade (Elisa and WB) tools will be cost-effective and useful for next large-scale epidemiological studies on Moroccan leishmaniasis animal reservoir.