Consensus PCR and Microarray for Diagnosis of the Genus Staphylococcus, Species, and Methicillin Resistance

S. Hamels, J.-L. Gala, S. Dufour, P. Vannuffel, N. Zammatteo, J. Remacle
2001 BioTechniques  
We propose the use of DNA microarray for the discrimination of homologous products after a single PCR amplification with consensus primers. The method was applied to Staphylococcus identification. The femA nucleotide sequences, which are phylogenetically conserved among the staphylococci , were first amplified using a consensus primer pair together with the mecA sequence, a molecular marker for methicillin resistance. Products were then identified on a glass array. The microarray contained five
more » ... rray contained five selective DNA capture probes for the simultaneous and differential identification of the five most clinically relevant staphylococcal species ( S. aureus , S. epidermidis, S. haemolyticus , S. hominis , and S. saprophyticus ), while a consensus capture probe could detect all femAsequences, allowing the identification of the genus Staphylococcus . The mecAsequence hybridized to a specific capture probe. The identification was univocal because only a single capture probe had to be present for each sequence to be identified. The hybridization and identification processes were completed in less than 2 h. Current results demonstrate that low-density microarrays are powerful multigenotypic post-PCR analyzers and could compete with conventional bacteria identification.
doi:10.2144/01316md04 pmid:11768666 fatcat:alq2zg7rxbftlbbyi4s372j7de