Recovery of long-chain lipopolysaccharides from liquid culture of Actinobacillus pleuropneumoniae (serotype 5) for ELISA serodiagnosis.
Actinobacillus pleuropneumoniae (serotype 5) long-chain lipopolysaccharides (LC-LPS) may be used as the antigen for ELISA serodiagnosis of pig pleuropneumonia. A method was developed in order to augment the recovery of this antigen compared to the previously described method. In liquid culture, A pleuropneumoniae was shown to produce 2.4 times more cells than in solid medium. LC-LPS could be recovered by phenol extraction of the crude extract. Only 1 additional phenol extraction was required to produce LC-LPS of the same quality as the one obtained with solid-medium-grown cells, as revealed by SDS-PAGE and ELISA using reference sera. From the same volume of crude extract, approximately 8 times more antigen was present in the aqueous phase of the phenol extraction.
Radacovici
S
S
Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Quebec, Canada.
Gottschalk
M
M
Dubreuil
J D
JD
D016977:Actinobacillus pleuropneumoniae / Q000276:immunology*; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D008070:Lipopolysaccharides / Q000302:isolation & purification*; D012698:Serologic Tests / Q000379:methods; D013552:Swine
D008070:Lipopolysaccharides