Hyperbranching Rolling Circle Amplification, An Improved Protocol for Discriminating Between Closely Related Fungal Species

Hyperbranching Rolling Circle Amplification (HRCA) is a technique derived from Rolling Circle Amplification (RCA) in which DNA polymerase replicates circularized oligonucleotide probes under isothermal conditions with either linear or geometric kinetics. Since its first introduction HRCA has been proven to be a robust DNA amplification technique for detecting pathogenic fungi and other applications, allowing rapid detection of nucleic-acid sequences with high specificity. Here we describe an improved protocol of HRCA, which is both specific and sensitive for detecting low copy numbers of template DNA. The test can be performed within one working day in routine molecular labs.