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

Document Type: Review Article

Authors

1 1Department of Biology, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Biology, Ferdowsi University of Mashhad, Mashhad, Iran; Research Department of Zoological Innovations, Institute of Applied Zoology, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

3 3Department of Parasitology and Mycology, Research Center for Cutaneous Leishmaniasis, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Parasitology and Mycology and Cancer Molecular Pathology Research Center, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Conventional methods for fungal identification in the clinical laboratory rely on morphological and
physiological tests. These tests often need several days or weeks to complete and are frequently
unspecific. Molecular identification mostly implies sequencing, which is relatively expensive and
time-consuming, as well as impractical for large numbers of isolates. The Rolling Circle
Amplification 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 for
the detection of pathogenic fungi. The specific probes are designed based on the differences in the
nucleotide regions of the target gene for the target species. The amplification product can be
visualized by agarose gel electrophoresis, but can also be visualized in gel-free systems using
fluorescence staining of the amplified product by SYBR Green in combination with a UV
transilluminator. Thus, the simplicity, sensitivity, robustness and low costs make RCA an attractive
technique for the reliable identification of sibling species and other closely related fungi.

Keywords