Abstract: The present invention provides a novel molecular method for the simultaneous identification and semi-quantification of multiple targeted biological entities from amongst a plurality. This invention discloses a method based on a multiplex nested amplification reaction in a single closed tube. The first amplification reaction relies on a set of large oligonucleotides for the amplification of common loci in all the targeted biological entities. The second nested amplification reaction relies on a set of short oligonucleotide primers that amplifies specific nucleotide sequences from all the amplicons previously produced in the first amplification reaction and generates an amplified product pattern capable of identifying each targeted biological entity. This method offers fast and accurate simultaneous identification of many targeted biological entities in any sample.