Molecular subtyping by genome and plasmid analysis ofCampylobacter jejuniserogroups O1 and O2 (Penner) from sporadic and outbreak cases of human diarrhoea

Abstract

SUMMARYRibosomal RNA gene patterns, randomly amplified polymorphic genomic DNA (RAPD) profiles and plasmid profiles were used to discriminate between 28 strains of Campylobacterjejuni serogroups 01 and 02 (Penner). Most isolates were biotype I (Lior). The strains were representative isolates from a UK school outbreak of enteritis (7 cases) and from 21 sporadic human cases of enteritis in 4 countries. The molecular techniques discriminated to various degrees between strains in each of the serogroups. The outbreak strains were homogeneous in most molecular features but a variety of types was detected amongst the isolates from the sporadic cases. Five groups of two or more strains with identical ribopatterns were identified and within each, strains from different patients were homogenous with respect to serogroup. RAPD profile typing based on numerical analysis generally matched ribotyping. Plasmid profiling overall gave least discrimination but was useful in separating some strains similar in other features. We concluded that optimal discrimination of C. jejuni could best be achieved using a combination of phenotypic and genotypic properties. Hae III ribotyping was the single most discriminatory and reproducible technique investigated. Several strains of C. jejuni from sporadic infections had similar molecular profiles which have potential for general typing purposes.

The application of genotyping techniques to the epidemiological analysis ofCampylobacter jejuni

“…Our results with the E3CJC2 probe, using a limited panel of strains, also found variability within these groups, particularly amongst strains of HS 1 (Table 2a). No genotype variation was detected amongst the HS 2/HL 4 strains examined in this study, contrary to the variability reported elsewhere [2,3,21]. However, our analysis is restricted to strains showing the defined stable antigenic associations predominating in human infections (e.g.…”

“…These three serogroups account for up to 80% of all serogroupable human isolates [17], and HaeIII ribotyping appears inadequate for subtyping this group. An earlier paper [3] reported far greater diversity in the Hae III ribotypes of HS 1 and HS 2 serogroup strains, but many of these isolates came from geographical areas outside the UK. We found Hae III ribotyping most discriminatory for serogroups which are less frequently isolated from humans ( Fig.…”

“…serogroup or phagetype determinants) [2][3][4][5]. High resolution genotyping methods can additionally evaluate the phylogenetic as well as epidemiologic relationships between strains [4,6].…”

Flagellin gene polymorphism analysis ofCampylobacter jejuniinfecting man and other hosts and comparison with biotyping and somatic antigen serotyping

“…Also there is molecular evidence that some strains within the same serogroup are genotypically diverse [7,29] so the discriminatory power of serotyping alone does not meet the requirements of a modern typing scheme. The aim of the present study was to develop a novel genotypic approach to categorizing isolates of C. jejuni that could be determined with relative simplicity and reproducibility.…”

Development and Application of a New Scheme for Typing Campylobacter jejuni and Campylobacter coli by PCR-Based Restriction Fragment Length Polymorphism Analysis

“…Genotyping methods are independent of expressed features and are therefore a better alternative for typing. Several genotyping methods have recently been developed, such as pulsed-field gel electrophoresis (11,12,40), amplified fragment length polymorphism (5,15), flagella gene PCR-restriction fragment length polymorphism (PCR-RFLP) (1,4,23,24,26), ribotyping (6,25,36), and random amplified polymorphic DNA analysis (7,10,30). These genotyping systems are more generally available and applicable than phenotypic methods.…”