Abstract: A method of identifying a plurality of microorganisms and their biomarkers for antibiotic resistance from a sample using Nucleic Acid amplification Technology (NAT) is provided. The method includes (i) extracting nucleic acid from a sample; (ii) preparing molecular biomarkers comprising a forward primer and a reverse primer for target genes of each of the plurality of microorganisms as a master mix; (iii) adding the extracted nucleic acid into the master mix to obtain a reaction mixture; (iv) performing a polymerase chain reaction (PCR) on the reaction mixture; (v) identifying a melt temperature for the amplicons; (vi) generating a melt curve for each target gene based on the identified melt temperatures and an amplification curve after the PCR amplification; and (vii) determining, using a Melt Curve analysis technique, Ct values, endpoint fluorescence level and melting profiles of the target genes by analyzing the amplification and melt curves to identify the plurality of microorganism present in the sample.