4.6 Article

One step for Legionella pneumophila detection in environmental samples by DNA-gold nanoparticle probe

Journal

JOURNAL OF APPLIED MICROBIOLOGY
Volume 125, Issue 5, Pages 1534-1540

Publisher

WILEY
DOI: 10.1111/jam.14047

Keywords

alternative tool; DNA probe; gold nanoparticle; Legionella pneumophila; rapid detection

Funding

  1. Khon Kaen University Research Grant [KKU-6100123]

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Aims To develop and evaluate a DNA-gold nanoparticle (DNA-AuNP) probe assay to detect Legionella pneumophila, which causes Legionnaires' disease, compared with the gold standard culture method. Methods and ResultsConclusionsGold nanoparticles (AuNPs) were conjugated with DNA probes to detect the mip gene of L. pneumophila. The DNA-AuNP probe assay was evaluated for its specificity, sensitivity and stability. The results showed that only L. pneumophila mixed with this probe resulted in a red solution that was easily detected by the naked eye, and the colour was stable when 10mmoll(-1) MgSO4 was added. The 100 Legionella isolates and 10 other bacteria led to 100% specificity. Compared with the culture method, our method showed a 100% negative predictive value, 100% sensitivity (kappa=087), and a detection limit of 45ng DNAl-1 with a 6-min response time for the 124 colonies suspected of being Legionella. The DNA-AuNP probe reagents were stable for more than 6months. The developed DNA-AuNP probe assay has good negative predictive value, sensitivity, rapidity and ease of use, which is helpful for ruling out negative samples. Significance and Impact of the StudyThe DNA-AuNP probe assay can detect the mip gene of L. pneumophila. Therefore, it may be an alternative method for screening colonies suspected of being L. pneumophila.

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