A new cost and time effective method for multilocus microsatellite typing (MLMT) studies: Application of Leishmania...

Molecular techniques are widely used in the field of parasitology to identify the genetic profile of the microbiological agents. Microsatellite typing studies are comprised of the amplification of polymorphic markers to analyze the fragment sizes using bioinformatics tools. Current methods need fluorescently labeled primers and size markers to obtain fragment peaks in ABI PRISM® systems and due to low discrimination power of gel-electrophoresis, it is not possible to differentiate primer-dimers from small fragments In the present study, we designed a new method for fragment analysis studies, which reduce the time by eliminating the classical PCR, the gel-electrophoresis and the preparation steps of fragment analysis. Ten previously studied Leishmania tropica strains and one Giemsa-stained slide were tested by new method and obtained fragment peaks were compared to the previous data obtained from ABI PRISM® system. Overall twelve makers were tested and the signal peak from each fragment was compared to classical ABI PRISM®-based fragment analysis and noted as identical. The new protocol is time saving, cost effective, and eliminates the human error comparing to classical MLMT analysis protocol. We believe that this method enables the easy detection of the fragment lengths without having bioinformatics experience and the obtained data can be easily shared with other laboratories.