sensitive colorimetric assay for identification of Acinetobacter baumannii using unmodified gold nanoparticles

AIMS: Acinetobacter baumannii is a global health problem, which threatens many healthcare settings. The current study aims to develop a detection assay for Ac. baumannii using unmodified gold nanoparticles (AuNPs). METHODS AND RESULTS: Fifty‐three Ac. baumannii clinical isolates were collected from...

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Bibliographic Details
Published in:	Journal of applied microbiology Vol. 117; no. 2; pp. 465 - 471
Main Authors:	Khalil, M.A.F, Azzazy, H.M.E, Attia, A.S, Hashem, A.G.M
Format:	Journal Article
Language:	English
Published:	Oxford Published for the Society for Applied Bacteriology by Blackwell Science 01-08-2014 Blackwell Oxford University Press
Subjects:	Acinetobacter baumannii Acinetobacter baumannii - genetics Acinetobacter baumannii - isolation & purification Bioassays Biological and medical sciences boiling citrates colorimetric assay colorimetry Colorimetry - methods detection limit DNA, Intergenic - chemistry Fundamental and applied biological sciences. Psychology Gold gold nanoparticles Gram-negative bacteria health services hospitals Humans in vitro detection intergenic DNA internal transcribed spacers Metal Nanoparticles - ultrastructure Microbiology nanogold Nanoparticles oligonucleotides pathogen identification PCR Polymerase Chain Reaction ribosomal RNA RNA, Ribosomal, 16S - genetics Applied microbiology Nanoparticle colorimetric assay gold nanoparticles Neisseriaceae Colorimetry Identification in vitro detection In vitro Polymerase chain reaction Acinetobacter baumannii Analysis method Bacteria Micrococcales Control method Detection PCR
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Summary:	AIMS: Acinetobacter baumannii is a global health problem, which threatens many healthcare settings. The current study aims to develop a detection assay for Ac. baumannii using unmodified gold nanoparticles (AuNPs). METHODS AND RESULTS: Fifty‐three Ac. 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 23S–16S rRNA intergenic spacer sequences (ITS). AuNPs were synthesized using citrate reduction method. Detection and optimization of Ac. baumannii amplicons using unmodified spherical AuNPs were performed using species‐specific DNA oligonucleotide. The nano‐gold assay was able to colorimetrically detect and distinguish Ac. baumannii from other Gram‐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 Ac. 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.
Bibliography:	http://dx.doi.org/10.1111/jam.12546 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1364-5072 1365-2672
DOI:	10.1111/jam.12546