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The Mycobacterium tuberculosis (MTB) shows different virulence and host infection range from other members of the M. tuberculosis complex (MTBC).Differential identification of MTB from MTBC is thus important in certain occasions.Many molecular diagnosis assays were developed that means the more advantages using molecular diagnosis than using conventional diagnosis in clinical specimens. In this study,we developed the diagnosis method that utilizes gold nanoparticles derivatized with thiol modified oligonucleotides that are designed to hybridize specifically complementaryDNA targets. Two pairs of gold nanoparticles probes, GP-IS6110 and GP-rv3618, aredesigned to specifically hybrid with target DNA of MTBC and MTB strains, respectively.