A New multiplex PCR have been developed in our laboratory using primer sets, aiming amplification of both D.N.A target fragments obtained in 18S RNA of commonly encountered fungi in human being and in Rhinosporidium seeberi using F1-fw/F2-rev (500 bp target) and Rhino-fw/ Rhino-rev (.177 bp target). This multiplex PCR has been found to be able to delect R. seeberi from clinical samples and differentiate it from other fungi. Furthermore, by this multiplex PCR, R. seeberi, phylogenitically appears to belong to a member of so called DRIPs clade of fish parasite not a cyanobacterium as claimed previously, by some workers.