Standardization of the Miniculture Technique for isolation of Leishmania spp. v1

Culture media are employed for the isolation of numerous biological agents causing diseases in humans and animals. The traditional culture method (TCM) is regarded as the gold standard in the diagnosis of Visceral and Tegumentary Leishmaniasis (VL / TL), as it is known that Leishmania spp. have a good growth on culture media used in TCM. The microculture and miniculture method consists in the use of tubes with a smaller volume, promoting a microaerophilic environment with high CO2 levels, which enhances the amastigote-to-promastigote differentiation. One of the disadvantages of culture methods is the possibility of contamination by bacteria and fungi, originating from the biological samples or the environment. It is important to ensure sterility during sample collection and processing. Therefore, the sample handling must be carried out with sterile material in a biological safety cabinet. This methodology requires trained personnel as well as sites adapted to biosafety standards for handling microorganisms class II. Although the microculture method is more sensitive than miniculture, the tubes used in this technique do not allow the parasite isolation and expansion. The miniculture tubes, like the traditional method, allows access to the isolate for post-culture analyses such as species characterization. Additionally, the miniculture showed better sensitivity compared to traditional methods and a shorter time to achieve positivity. Furthermore, miniculture is easy to implement, using the same equipment and materials as traditional culture but in smaller volumes, which can lead to cost reduction.