Affiliation:
1. Department of Bacteriology, Institute for Animal Science and Health, 8200 AB Lelystad, The Netherlands,1 and
2. Institute of Medical Microbiology and Hygiene, Johannes Gutenberg University, 55101 Mainz, Germany2
Abstract
ABSTRACT
For epidemiological studies of
Campylobacter
infections, molecular typing methods that can differentiate campylobacters at the strain level are needed. In this study we used a recently developed genotyping method, amplified fragment length polymorphism (AFLP), which is based on selective amplification of restriction fragments of chromosomal DNA, for genetic typing of
Campylobacter jejuni
and
Campylobacter coli
strains derived from humans and poultry. We developed an automated AFLP fingerprinting method in which restriction endonucleases
Hin
dIII and
Hha
I were used in combination with one set of selective PCR primers. This method resulted in evenly distributed band patterns for amplified fragments ranging from 50 to 500 bp long. The discriminatory power of AFLP was assessed with a
C. jejuni
strain, an isogenic flagellin mutant, and distinct
C. jejuni
strains having known pulsed-field gel electrophoresis and
fla
PCR-restriction fragment length polymorphism genotypes. Unrelated
C. jejuni
strains produced heterogeneous patterns, whereas genetically related strains produced similar AFLP patterns. Twenty-five
Campylobacter
strains obtained from poultry farms in The Netherlands grouped in three
C. jejuni
clusters that were separate from a
C. coli
cluster. The band patterns of 10
C. jejuni
strains isolated from humans were heterogeneous, and most of these strains grouped with poultry strains. Our results show that AFLP analysis can distinguish genetically unrelated strains from genetically related strains of
Campylobacter
species. However, desirable genetically related strains can be differentiated by using other genotyping methods. We concluded that automated AFLP analysis is an attractive tool which can be used as a primary method for subtyping large numbers of
Campylobacter
strains and is extremely useful for epidemiological investigations.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
135 articles.
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