This document provides updated results on the second Leishmania External Quality Assessment (EQA), established to be performed among partners for optimization activity to achieve reliable results in molecular diagnosis in sand fly samples collected during field activities (Del. 2.1). Reference laboratories were Istituto Superiore di Sanità (ISS)/ Université d'aix Marseille (AMU) for phleboviruses and ISS/ Institut National de la Santé et de la Recherche Médicale (INSERM) for Leishmania detection and identification. Also, IFAT EQA results are provided. Information on the percentage of analysed pools per site, and pathogens screening conducted in 10 countries by 9 partners are included.