Open AccessA dual approach employing MALDI ‐ TOF MS and real‐time PCR for fast species identification within the E nterobacter cloacae complex
Author(s) -
Pavlovic Melanie,
Konrad Regina,
Iwobi Azuka N.,
Sing Andreas,
Busch Ulrich,
Huber Ingrid
Publication year - 2012
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2011.02479.x
Subject(s) - enterobacter cloacae , enterobacter , enterobacteriaceae , biology , microbiology and biotechnology , matrix assisted laser desorption/ionization , chemistry , gene , biochemistry , escherichia coli , desorption , organic chemistry , adsorption
A real‐time PCR procedure targeting the gene of the molecular cochaperon DnaJ ( dnaJ ) was developed for specific detection of strains belonging to the E nterobacter cloacae group. The inclusivity and exclusivity of the real‐time PCR assay were assessed with seven reference strains of E . cloacae , 12 other E nterobacter species and 41 non‐ E nterobacter strains. Inclusivity as well as exclusivity of the duplex real‐time PCR was 100%. In contrast, resolution of matrix‐assisted laser desorption ionization‐time of flight mass spectrometry ( MALDI ‐ TOF MS ) was inadequate for delineation of E nterobacter asburiae , E nterobacter hormaechei , E nterobacter kobei and E nterobacter ludwigii from E. cloacae . Eleven of 56 (20%) clinical isolates of the E . cloacae group could not be clearly identified as a certain species using MALDI ‐ TOF MS . In summary, the combination of MALDI ‐ TOF MS with the E . cloacae ‐specific duplex real‐time PCR is an appropriate method for identification of the six species of the E . cloacae complex.