Development of an Immunoassay for Rapid Detection of Ganglioside GM1 Mimicry in Campylobacter jejuniStrains

摘要

Mimicry of peripheral nerve gangliosides by Campylobacter jejuni lipopolysaccharides (LPSs) has been proposed to induce cross-reacting antiganglioside antibodies in Guillain-Barré syndrome (GBS). Because current methods for LPS characterization are labor-intensive and inhibit the screening of large numbers of strains, a rapid GM₁ epitope screening assay was developed. Biomass from two agar plates of confluent growth yielded sufficient LPS using a novel phenol-water and ether extraction procedure. Extracts of LPS were reacted with cholera toxin (GM₁ ligand), peanut agglutinin (Galβ1→3GalNAc ligand), and anti-GM₁ antibodies. After the assay was validated, 12 of 59 (20%) C. jejuni serostrains, including four serotypes that have not previously been associated with GBS, reacted with two or more anti-GM₁ ganglioside reagents. Subsequently, LPS extracts from 5 of 7 (71%) C. jejuniisolates and 2 of 3 (67%) C. jejuni culture collection strains bore GM₁ structures. Overall, the assay system was reliable, efficient, and reproducible and may be adapted for large-scale epidemiological studies.