Development of loop-mediated isothermal amplification for rapid detection of sporotrichosis caused by Sporothrix schenckii

Background and Aim: Sporothrix schenckii is the causative agent of sporotrichosis, which most commonly causes lymphocutaneous infections in immunocompromised hosts. This pathogen infects dogs, cats, cattle, buffaloes can potentially infect humans. Diagnosis by fungal culture lengthy, although there are several clinical diagnoses molecular methods, these complicated time-consuming for veterinarians. study aimed to develop a visual diagnostic assay that less be used veterinarians screen sporotrichosis. Materials Methods: To loop-mediated isothermal amplification (LAMP) primers specific fragments 18S rRNA gene S. were designed. Then, time temperature optimized successfully achieve LAMP. Ten-fold serial dilutions DNA determine detection limit using both LAMP nested polymerase chain reaction (nPCR) assays. Results: The optimal conditions incubation at 73°C 30 min. Agarose gel electrophoresis revealed ladder-like pattern product, sky-blue color indicated positive result. A comparison with nPCR it was 10 times more sensitive than nPCR, pg. use heat box compared thermocycler gave same results. Conclusion: Loop-mediated gives good results may represent future alternative tool screening pathogens before conventional cultures received. However, this method should further studied clarify its samples. Keywords: amplification, detection, rapid schenckii, detection.