High density whole genome fingerprinting of methicillin-resistant and -susceptible strains of Staphylococcus aureus in search of phenotype-specific molecular determinants.
AFLP is a selective restriction fragment amplification method generating DNA fingerprints for microbial isolates. We present high-throughput AFLP (htAFLP) to characterize molecular markers associated with bacterial phenotypes. Methicillin-resistant and -susceptible isolates of Staphylococcus aureus have been used for this model study in conjunction with the available S. aureus genome sequences. This facilitates the calculation of theoretical AFLP fingerprints, comparison of these fingerprints with genuine experimental fingerprints, and the subsequent identification of polymorphic AFLP markers without sequence analysis. Analysis of 46 MRSA and 46 MSSA strains by 39 different AFLP reactions generated more than 2500 fragments per strain and an overall number of 6180 scorable markers within all strains. We successfully identify MRSA specific markers and elaborate on the general applicability of the htAFLP approach. This method can be applied to any microbial species for which at least one full-genome sequence is available.
Savelkoul
Paul H M
PH
VU University Medical Center, Department of Medical Microbiology & Infection Control, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands. p.savelkoul@vumc.nl
Melles
Damian C
DC
Buffing
Nico
N
Gorkink
Roy
R
Simons
Guus
G
van Belkum
Alex
A
D015373:Bacterial Typing Techniques; D016172:DNA Fingerprinting / Q000379:methods; D004269:DNA, Bacterial; D024881:Drug Resistance, Bacterial; D005819:Genetic Markers; D016680:Genome, Bacterial; D008712:Methicillin / Q000494:pharmacology; D016106:Methicillin Resistance / Q000235:genetics*; D010641:Phenotype; D013203:Staphylococcal Infections / Q000453:epidemiology / Q000382:microbiology; D013211:Staphylococcus aureus / Q000145:classification / Q000187:drug effects / Q000235:genetics*
D004269:DNA, Bacterial; D005819:Genetic Markers; D008712:Methicillin