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Subtyping of Campylobacter jejuni ssp. doylei Isolates Using Mass Spectrometry-based PhyloProteomics (MSPP)
Published on: October 30, 2016
Identification of DNA sequence variation in Campylobacter jejuni strains associated with the Guillain-Barré syndrome
Peggy C R Godschalk1, Mathijs P Bergman, Raymond F J Gorkink
1Department of Medical Microbiology & Infectious Diseases, Erasmus MC--University Medical Center Rotterdam, Dr, Molewaterplein 40, 3015 GD Rotterdam, The Netherlands. p.godschalk@erasmusmc.nl
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.
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.
Insights
Campylobacter jejuni infections can trigger Guillain-Barré syndrome (GBS) through molecular mimicry. A new htAFLP method identified specific bacterial markers in LOS biosynthesis genes linked to GBS.
Area of Science:
- Microbiology
- Immunology
- Genetics
Background:
- Campylobacter jejuni is a leading cause of infections preceding Guillain-Barré syndrome (GBS) and Miller Fisher syndrome (MFS).
- The mechanism likely involves molecular mimicry between human gangliosides and bacterial lipo-oligosaccharides (LOS).
- Identifying specific bacterial markers is crucial for understanding GBS pathogenesis.
Purpose of the Study:
- To develop and validate a high-throughput AFLP (htAFLP) method for detecting DNA polymorphisms in C. jejuni.
- To identify C. jejuni markers specifically associated with Guillain-Barré syndrome (GBS) and Miller Fisher syndrome (MFS).
Main Methods:
- A novel htAFLP approach was employed to analyze DNA polymorphism in C. jejuni strains.
- PCR-RFLP and DNA sequencing were used to validate htAFLP findings.
- Strains associated with GBS/MFS and enteritis were analyzed for specific markers.
Main Results:
- HtAFLP generated thousands of markers, revealing polymorphisms between C. jejuni isolates.
- Comparison with GBS-associated strains identified 23 potential markers, with 17 sequenced.
- Three markers in LOS biosynthesis genes (cj1136, cj1138, cj1139c) were significantly associated with GBS.
Conclusions:
- Bacterial GBS markers are few and located in LOS biosynthesis genes, supporting the LOS mimicry hypothesis.
- The htAFLP technique demonstrates high reproducibility and resolution for identifying bacterial disease markers.
- This method aids in detecting and identifying putative bacterial markers for GBS and related neuropathies.
