Rolling Circle Amplification (RCA): an approach for quick detection and identification of fungal species

Conventional methods for fungal identification in the clinical laboratory rely on morphological andphysiological tests. These tests often need several days or weeks to complete and are frequentlyunspecific. Molecular identification mostly implies sequencing, which is relatively expensive andtime-consuming, as well as impractical for large numbers of isolates. The Rolling CircleAmplification approach, known as RCA, is a quick, critical and economic method for fungal species'identification. Despite its high speed, this method is highly sensitive, and it has been widely used forthe detection of pathogenic fungi. The specific probes are designed based on the differences in thenucleotide regions of the target gene for the target species. The amplification product can bevisualized by agarose gel electrophoresis, but can also be visualized in gel-free systems usingfluorescence staining of the amplified product by SYBR Green in combination with a UVtransilluminator. Thus, the simplicity, sensitivity, robustness and low costs make RCA an attractivetechnique for the reliable identification of sibling species and other closely related fungi.