Identification of DNA sequence variation in Campylobacter jejuni strains associated with the Guillain-Barré syndrome by high-throughput AFLP analysis.
BACKGROUND
Campylobacter jejuni is the predominant cause of antecedent infection in post-infectious neuropathies such as the Guillain-Barré (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.


CONCLUSIONS
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.
Godschalk
Peggy C R
PC
Department of Medical Microbiology & Infectious Diseases, Erasmus MC--University Medical Center Rotterdam, Dr, Molewaterplein 40, 3015 GD Rotterdam, The Netherlands. p.godschalk@erasmusmc.nl
Bergman
Mathijs P
MP
Gorkink
Raymond F J
RF
Simons
Guus
G
van den Braak
Nicole
N
Lastovica
Albert J
AJ
Endtz
Hubert P
HP
Verbrugh
Henri A
HA
van Belkum
Alex
A
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002169:Campylobacter Infections / Q000382:microbiology*; D016123:Campylobacter jejuni / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D005732:Gangliosides; D005819:Genetic Markers; D014644:Genetic Variation; D020275:Guillain-Barre Syndrome / Q000382:microbiology*; D006801:Humans; D008070:Lipopolysaccharides / Q000096:biosynthesis; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005732:Gangliosides; D005819:Genetic Markers; D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides