Use of fluorescent amplified fragment length polymorphism (fAFLP) to characterise methicillin-resistant Staphylococcus aureus

• Published in: Journal of microbiological methods 1999-07, Vol.37 (1), p.7-15
• Main Authors: Hookey, J.V, Edwards, V, Patel, S, Richardson, J.F, Cookson, B.D
• Format: Article
• Language: English
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Coagulase - genetics; Electrophoresis, Gel, Pulsed-Field - methods; Fluorescent amplified fragment length polymorphism; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genotype; Methicillin Resistance - genetics; Microbiology; Phenotype; Polymerase Chain Reaction - methods; Polymorphism, Restriction Fragment Length; Staph aureus; Staphylococcus aureus; Staphylococcus aureus - classification; Staphylococcus aureus - drug effects; Staphylococcus aureus - genetics
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/S0167-7012(99)00023-8

The new PCR-based genotyping technique, fluorescent amplified fragment length polymorphism (fAFLP), was compared for discriminatory power and reproducibility with standard phenotypic methods, a coagulase gene ( coa) restriction fragment length polymorphism (RFLP) method and pulsed-field gel electrophoresis (PFGE), in typing 34 isolates and four reference strains of methicillin-resistant Staphylococcus aureus (MRSA). The fAFLP showed from 40 to 75 fragments, 50 to 450 base pairs (bp) in size. Based on replicate studies, the isolates were judged indistinguishable when their fAFLP pattern was >93.7% similar. Only two of the isolates were indistinguishable by this criterion. Thirty-one MRSA fell into four major fAFLP groups (1, 2, 3 and 4) at the level of >79.9% similarity. Three other isolates and an EMRSA-16 strain fell outside these major groups. Within both fAFLP groups 1 and 2, two subgroups, A and B, could be identified at ∼82.0% similarity. While most isolates within group 1 could also be separated by their phenotypic and coagulase gene ( coa) RFLP pattern, all the isolates within fAFLP groups 2A and 2B were identical on the basis of these characters. The MRSA within fAFLP groups 3 and 4 were heterogeneous by their phenotypic characteristics and coa gene RFLP patterns. fAFLP was reproducible and distinguished between MRSA isolates that appeared identical by other methods. It is likely to contribute to the epidemiological analysis of outbreaks of MRSA infection.