Development and Evaluation of a Molecular Diagnostic Method for Rapid Detection of Histoplasma capsulatum var. farciminosum, the Causative Agent of Epizootic Lymphangitis, in Equine Clinical Samples

摘要: Histoplasma capsulatum var. farciminosum, the causative agent of epizootic lymphangitis (EZL), is endemic in parts Africa. Diagnosis based on clinical signs and microscopy lacks specificity a barrier to further understanding this neglected disease. Here, nested PCR method targeting internal transcribed spacer (ITS) region rRNA operon was validated for application equine samples. Twenty-nine horses with EZL from different climatic regions Ethiopia were clinically examined. Blood samples aspirates pus cutaneous nodules taken, along blood 20 no lesions. Among 29 suspected cases EZL, H. farciminosum confirmed by extraction DNA 25 17 horses, respectively. Positive results also obtained heat-inactivated (24 horses) (23 spotted onto Whatman FTA cards. Two positive among that did not exhibit EZL. These are first reports direct detection at high frequency exhibiting The outperformed conventional microscopic diagnosis, as characteristic yeast cells could be observed only 14 presence sequencing cloned products, while alignment ITS amplicons showed very little sequence variation, there preliminary single nucleotide polymorphism-based evidence existence two subgroups This molecular diagnostic now permits investigation epidemiology