Exploration of amplified fragment length polymorphism analysis as an effective tool for discriminating pathogenic strains of bacteria

Abstract

Exploration of amplified fragment length polymorphism analysis as an effective tool for discriminating pathogenic strains of bacteria K Weinbrecht,1 A Taylor,2 C Beaumann,3 RW Allen11Department of Forensic Sciences, Center for Health Sciences, Oklahoma State University, Tulsa, OK; 2Centers for Disease Control, Atlanta, GA; 3PharmaNet-i3, Indianapolis, IN, USABackground: The detrimental effect pathogens used as bioweapons could have on the US is sufficient to warrant extensive efforts to establish preparedness to prevent, and, if necessary, respond to such an event. The objective of this research was to adapt and refine amplified fragment length polymorphism (AFLP) analysis for DNA profiling of microbial strains for use in forensic and clinical case work.Methods: Included in this study were multiple strains of Pseudomonas syringae that were restricted in their respective host ranges (ie, the pathovars maculicola and tomato), Pseudomonas aeruginosa, Serratia marcescens, and Staphylococcus aureus. AFLP profiles were reduced to numeric haplotype codes that conveyed the size characteristics of the profiles.Results: The AFLP assay exhibited over 97% reproducibility and every analyzed strain produced a unique haplotype code. Conserved areas of similarity were identified in haplotype codes of closely related strains of the same species, indicating that AFLP may also be a useful species identification tool, a possibility supported by cluster analysis of AFLP codesConclusion: The results of this study demonstrate that AFLP technology is sufficiently reproducible, powerful, and reliable for use as a broadly effective molecular screening tool in microbial forensics.Keywords: amplified fragment length polymorphism, pathogenic strains, bacteria