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A sensitive colorimetric assay for identification of A cinetobacter baumannii using unmodified gold nanoparticles
Author(s) -
Khalil M.A.F.,
Azzazy H.M.E.,
Attia A.S.,
Hashem A.G.M.
Publication year - 2014
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12546
Subject(s) - acinetobacter baumannii , colloidal gold , microbiology and biotechnology , detection limit , amplicon , dna extraction , oligonucleotide , gold standard (test) , biology , bacteria , polymerase chain reaction , chromatography , dna , chemistry , medicine , nanoparticle , biochemistry , nanotechnology , gene , materials science , genetics , pseudomonas aeruginosa
Aims A cinetobacter baumannii is a global health problem, which threatens many healthcare settings. The current study aims to develop a detection assay for A c. baumannii using unmodified gold nanoparticles ( A u NP s). Methods and Results Fifty‐three A c. baumannii clinical isolates were collected from Egyptian hospitals. Bacterial isolation and biochemical identification of isolates were carried out followed by DNA extraction using boiling method and PCR amplification of the 23 S –16 S rRNA intergenic spacer sequences ( ITS ). Au NP s were synthesized using citrate reduction method. Detection and optimization of A c. baumannii amplicons using unmodified spherical A u NP s were performed using species‐specific DNA oligonucleotide. The nano‐gold assay was able to colorimetrically detect and distinguish A c. baumannii from other G ram‐negative bacteria. The turnaround time of the assay is about 2 h including sample treatment and amplification. The assay detection limit is 0·8125 ng of DNA . Conclusions The developed colorimetric assay is sensitive, fast and reliable and can be used for identification of A c. baumannii . Significance and Impact of the Study There is a need to develop robust, rapid, and specific methods for detection of Ac. baumannii isolated from clinical specimens. The developed nanogold assay prototype allows sensitive, specific and rapid detection of amplified DNA of A . baumannii and represents a reliable diagnostic tool to aid routine laboratory identification of this pathogen.