A technique for multiplex detection of pathogens based on padlock probes and Luminex technology has been developed. Here detection data from a panel of pathogenic fungi is presented. Most nucleic acid based techniques in use for clinical pathogen detection (diagnosis) can reveal one to a few pathogens in each test. Techniques revealing several pathogens at a time are often problematic due to inherent difficulties with multiplex PCR. The padlock probe (PLP) based detection method, presented here, avoids such problems and has potential for developing highly multiplexed assays. A PLP is an oligonucleotide designed to hybridize to a target sequence so that the 5’ and 3’ end of the probe meet with only a nick separating them. A ligase can close the nick, forming a closed DNA circle of the probe. In the case presented here the internal parts of the probe (the parts not participating in target hybridization) have a general PCR primer target region and a DNA address region (tag region). General PCR primers, that are labelled, can amplify circularized probes and the products can be detected in a Luminex™ machine via the address tags.