Rapid identification of bacterial pathogens using a PCR- and microarray-based assay |
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Authors: | Anna-Kaarina Järvinen Sanna Laakso Pasi Piiparinen Anne Aittakorpi Merja Lindfors Laura Huopaniemi Heli Piiparinen Minna Mäki |
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Institution: | (1) Mobidiag Ltd, 00290 Helsinki, Finland |
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Abstract: | Background During the course of a bacterial infection, the rapid identification of the causative agent(s) is necessary for the determination
of effective treatment options. We have developed a method based on a modified broad-range PCR and an oligonucleotide microarray
for the simultaneous detection and identification of 12 bacterial pathogens at the species level. The broad-range PCR primer
mixture was designed using conserved regions of the bacterial topoisomerase genes gyrB and parE. The primer design allowed the use of a novel DNA amplification method, which produced labeled, single-stranded DNA suitable
for microarray hybridization. The probes on the microarray were designed from the alignments of species- or genus-specific
variable regions of the gyrB and parE genes flanked by the primers. We included mecA-specific primers and probes in the same assay to indicate the presence of methicillin resistance in the bacterial species.
The feasibility of this assay in routine diagnostic testing was evaluated using 146 blood culture positive and 40 blood culture
negative samples. |
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