BackgroundDue to the advantages of molecular methods over biochemical methods, the use of molecular methods for diagnosing nosocomial infections such asPseudomonascan be an appropriate and rapid way to choose the right diagnosis and treatment of infection and prevent further complications caused by the infection. The present article provides a description of the development of a nanoparticle-based detection technique for sensitive and specific deoxyribonucleic acid-based diagnostic ofPseudomonas aeruginosa. Specific thiolated oligonucleotide probes for one of the hypervariable regions of the16S rDNAgene were designed and applied for colorimetric detection of the bacteria.ResultsThe results of gold nanoprobe-nucleic sequence amplification indicated the probe attached to gold nanoparticles in the presence of the target deoxyribonucleic acid. It caused aggregation of gold nanoparticles in the form of connected networks resulting in color change and indicating the presence of the target molecule in the sample, which could be observed by the naked eye. In addition, the wavelength of gold nanoparticles changed from 524 to 558 nm. Multiplex polymerase chain reactions were performed using four specific genes ofPseudomonas aeruginosa(oprL,oprI,toxA, and16S rDNA). The sensitivity and specificity of the two techniques were assessed. According to the observations, the specificity of both techniques was 100%, and the sensitivity was 0.5 ng/μL and 0.01 ng/μL of genomic deoxyribonucleic acid for multiplex polymerase chain reaction and colorimetric assay, respectively.ConclusionsThe sensitivity of colorimetric detection was about 50 times higher than the polymerase chain reaction using the16SrDNAgene. The results of our study proved to be highly specific with potential use for early detection ofPseudomonas aeruginosa.