Multiplex PCR to detect the genes encoding naturally occurring oxacillinases in Acinetobacter spp

Kamolvit, Witchuda, Higgins, Paul G., Paterson, David L. and Seifert, Harald (2014) Multiplex PCR to detect the genes encoding naturally occurring oxacillinases in Acinetobacter spp. Journal of Antimicrobial Chemotherapy, 69 4: 959-963. doi:10.1093/jac/dkt480


Author Kamolvit, Witchuda
Higgins, Paul G.
Paterson, David L.
Seifert, Harald
Title Multiplex PCR to detect the genes encoding naturally occurring oxacillinases in Acinetobacter spp
Formatted title
Multiplex PCR to detect the genes encoding naturally occurring oxacillinases in Acinetobacter spp 
Journal name Journal of Antimicrobial Chemotherapy   Check publisher's open access policy
ISSN 0305-7453
1460-2091
Publication date 2014-01-01
Year available 2013
Sub-type Article (original research)
DOI 10.1093/jac/dkt480
Volume 69
Issue 4
Start page 959
End page 963
Total pages 5
Place of publication Oxford, United Kingdom
Publisher Oxford University Press
Language eng
Formatted abstract
Objectives: Bacteria of the genus Acinetobacter are increasingly being isolated in hospitals and are recognized as emerging nosocomial pathogens. Species identification is difficult and there is a need for simple molecular methods to differentiate between the species. Naturally occurring oxacillinase genes (blaOXA) have been identified in several Acinetobacter species and their detection by PCR can aid in species identification. The aim of this study was to develop a multiplex PCR to identify intrinsic blaOXA genes (i.e. blaOXA-134-like, blaOXA-211-like, blaOXA-213-like, blaOXA-214-like and blaOXA-228-like) from Acinetobacter spp. for use as a tool for rapid species identification.

Methods: Primers were designed to selectively amplify internal fragments of intrinsic blaOXA from Acinetobacter lwoffii/Acinetobacter schindleri (blaOXA-134-like), Acinetobacter johnsonii (blaOXA-211-like), Acinetobacter calcoaceticus (blaOXA-213-like), Acinetobacter haemolyticus (blaOXA-214-like) and Acinetobacter bereziniae (blaOXA-228-like). Multiplex PCR was performed in a total of 100 Acinetobacter isolates. Flanking primers were designed for each blaOXA subgroup and products were sequenced.

Results: All A. lwoffii, A. schindleri, A. johnsonii, A. calcoaceticus, A. haemolyticus and A. bereziniae isolates were positive for their species-specific amplicons while other Acinetobacter species were negative. Thirty blaOXA novel variants were identified; the majority of these (21/30) were from A. calcoaceticus. ISAba11 was found upstream of blaOXA-214 in four A. haemolyticus isolates, but was not associated with carbapenem resistance.

Conclusions: This multiplex PCR specifically detected each of the five different blaOXA subgroups. Therefore, this method has the potential to aid in the identification of these species and monitor the spread of these genes into other Acinetobacter species.
Keyword Carbapenemases
Intrinsic OXA
Species identification
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: UQ Centre for Clinical Research Publications
Official 2014 Collection
 
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