Identification of DNA sequence variation in Campylobacter jejuni strains associated with the Guillain-Barre syndrome by high-throughput AFLP analysis.
BACKGROUND: Campylobacter jejuni is the predominant cause of antecedent infection in post-infectious neuropathies such as the Guillain-Barre (GBS) and Miller Fisher syndromes (MFS). GBS and MFS are probably induced by molecular mimicry between human gangliosides and bacterial lipo-oligosaccharides (LOS). This study describes a new C. jejuni-specific high-throughput AFLP (htAFLP) approach for detection and identification of DNA polymorphism, in general, and of putative GBS/MFS-markers, in particular. RESULTS: We compared 6 different isolates of the "genome strain" NCTC 11168 obtained from different laboratories. HtAFLP analysis generated approximately 3000 markers per stain, 19 of which were polymorphic. The DNA polymorphisms could not be confirmed by PCR-RFLP analysis, suggesting a baseline level of 0.6% AFLP artefacts. Comparison of NCTC 11168 with 4 GBS-associated strains revealed 23 potentially GBS-specific markers, 17 of which were identified by DNA sequencing. A collection of 27 GBS/MFS-associated and 17 enteritis control strains was analyzed with PCR-RFLP tests based on 11 of these markers. We identified 3 markers, located in the LOS biosynthesis genes cj1136, cj1138 and cj1139c, that were significantly associated with GBS (P = 0.024, P = 0.047 and P < 0.001, respectively). HtAFLP analysis of 13 highly clonal South African GBS/MFS-associated and enteritis control strains did not reveal GBS-specific markers. CONCLUSION: This study shows that bacterial GBS markers are limited in number and located in the LOS biosynthesis genes, which corroborates the current consensus that LOS mimicry may be the prime etiologic determinant of GBS. Furthermore, our results demonstrate that htAFLP, with its high reproducibility and resolution, is an effective technique for the detection and subsequent identification of putative bacterial disease markers.
|Keywords||*Polymorphism, Restriction Fragment Length, *Variation (Genetics), Bacterial Proteins/genetics, Base Sequence, Campylobacter Infections/*microbiology, Campylobacter jejuni/*genetics, DNA, Bacterial/genetics, Gangliosides, Genetic Markers, Guillain-Barre Syndrome/*microbiology, Humans, Lipopolysaccharides/biosynthesis, Molecular Mimicry, Molecular Sequence Data, Polymerase Chain Reaction/*methods, Polymorphism, Genetic|
|Persistent URL||dx.doi.org/10.1186/1471-2180-6-32, hdl.handle.net/1765/13996|
Godschalk, P.C.R., Bergman, M.P., Gorkink, R.F.J., Simons, G., van den Braak, N.P.W.C.J., Lastovica, A.J., … van Belkum, A.F.. (2006). Identification of DNA sequence variation in Campylobacter jejuni strains associated with the Guillain-Barre syndrome by high-throughput AFLP analysis.. B M C Microbiology, 6. doi:10.1186/1471-2180-6-32