Microtiter assay for detecting Campylobacter spp. and Helicobacter pylori with surface gangliosides which bind cholera toxin.
Campylobacter jejuni with Gm1 ganglioside in the core of its lipopolysaccharide has been associated with Guillain-Barré syndrome. Since this epitope may be of considerable pathophysiologic importance and since this ganglioside binds cholera toxin, a rapid screening assay to detect bacteria that bind cholera toxin as an indication of Gm1 on their surfaces was developed. In the assay, bacterial lawns were grown on agar plates, harvested with phosphate-buffered saline, boiled, and incubated with a standard concentration of cholera B subunit. Preparations from strains with Gm1 were observed to inhibit the binding of cholera B subunit to Gm1 in a microtiter enzyme-linked immunosorbent assay. By using this assay with two groups of strains, 37 positive strains were detected among the 197 tested. Species with positive isolates included C. jejuni, Campylobacter coli, and Helicobacter pylori. The assay is capable of testing large numbers of isolates and should prove useful in future clinical and epidemiological studies of bacteria with this epitope.
Sack
D A
DA
Department of International Health, Johns Hopkins University, Baltimore, Maryland 21205, USA. dsack@jhsph.edu
Lastovica
A J
AJ
Chang
S H
SH
Pazzaglia
G
G
D001431:Bacteriological Techniques; D001667:Binding, Competitive; D002167:Campylobacter / Q000302:isolation & purification* / Q000378:metabolism; D002169:Campylobacter Infections / Q000175:diagnosis*; D002772:Cholera Toxin / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005677:G(M1) Ganglioside / Q000378:metabolism*; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans
D005677:G(M1) Ganglioside; D002772:Cholera Toxin