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The WHO multicenter study on Listeria monocytogenes subtyping: random amplification of polymorphic DNA (RAPD)
K Wernars1, P Boerlin, A Audurier
1Microbiological Laboratory for Health Protection, National Institute of Public Health and the Environment, Bilthoven, Netherlands. Karel.Wernars@rivm.nl
International Journal of Food Microbiology
|October 1, 1996
Summary
The random amplification of polymorphic DNA (RAPD) technique showed variable reproducibility in Listeria monocytogenes subtyping across laboratories. Further optimization is needed for reliable epidemiological analysis using this molecular method.
Area of Science:
- Microbiology
- Molecular Biology
- Epidemiology
Background:
- Listeria monocytogenes is a significant foodborne pathogen requiring accurate subtyping for outbreak investigations.
- Standardized molecular subtyping methods are crucial for public health surveillance.
- The random amplification of polymorphic DNA (RAPD) technique is a potential tool for bacterial fingerprinting.
Purpose of the Study:
- To evaluate the reproducibility and discriminatory power of the random amplification of polymorphic DNA (RAPD) technique for Listeria monocytogenes subtyping.
- To assess the performance of a standardized RAPD protocol across multiple participating laboratories.
- To determine the suitability of RAPD for epidemiological investigations of Listeria monocytogenes.
Main Methods:
- A World Health Organization (WHO) multicenter study involving six participants.
- Standardized random amplification of polymorphic DNA (RAPD) protocol applied to 80 Listeria monocytogenes strains.
- Analysis included epidemiologically linked isolates and duplicate strains using three different 10-mer primers.
Main Results:
- Median reproducibility of RAPD results across participants was 86.5%, with significant variation (0-100%).
- Inconsistent results were primarily linked to one specific primer.
- The number of homogeneous epidemiological groups identified varied widely among participants (median 16), with some groups showing inhomogeneity.
- Overall correlation between participants' results ranged from 32% to 85%.
Conclusions:
- The random amplification of polymorphic DNA (RAPD) technique, as evaluated, exhibits limitations in reproducibility and inter-laboratory consistency for Listeria monocytogenes subtyping.
- Variability in results suggests challenges in standardizing RAPD protocols for reliable epidemiological surveillance.
- Further refinement and validation of RAPD methods are necessary to enhance its utility in public health.