Detection of Campylobacter jejuni in food and poultry viscera using immunomagnetic separation and microtitre hybridization.
Thermophillic Campylobacter and Camp. jejuni were detected from samples of chicken liver, gall bladder, muscle and contaminated milk and chicken meat after an enrichment step by using immunomagnetic capture of cells with monoclonal antibody against a specific outer membrane protein of thermophilic Campylobacter. The detection of captured cells was achieved using two different hybridization methods. In one of the methods, the captured cells were lysed by guanidine isothiocyanate and the 23S rRNA was reacted with a microtitre plate-immobilized rDNA probe specific for thermophilic Campylobacter. In the other method, the captured cells were subjected to lysis by ultrasonication and the genomic DNA reacted with a microtitre plate-immobilized RNA probe specific for Camp.jejuni. Detection of the RNA-DNA hybrids formed in the wells was carried out using a monoclonal anti-RNA-DNA hybrid antibody.
Lamoureux
M
M
Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada.
MacKay
A
A
Messier
S
S
Fliss
I
I
Blais
B W
BW
Holley
R A
RA
Simard
R E
RE
D000818:Animals; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000032:analysis / Q000276:immunology; D016123:Campylobacter jejuni / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis / Q000276:immunology; D005704:Gallbladder / Q000382:microbiology; D018189:Immunomagnetic Separation; D008099:Liver / Q000382:microbiology; D008892:Milk / Q000382:microbiology*; D018482:Muscle, Skeletal / Q000382:microbiology; D009693:Nucleic Acid Hybridization; D011202:Poultry Products / Q000382:microbiology*; D012329:RNA, Bacterial / Q000032:analysis / Q000276:immunology
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D012329:RNA, Bacterial