Microtitre plate hybridization system for detection of thermophilic Campylobacter rRNA.
A microtitre plate nucleic acid probe hybridization system was developed for the detection of ribosomal RNA from thermophilic Campylobacter (Camp. jejuni, Camp. coli, Camp. lari and Camp. upsaliensis). A specific DNA probe obtained by amplification of 23S rRNA sequences using the polymerase chain reaction technique was immobilized on a microtitre plate, and used for hybridization with target 23S rRNA from cell lysates. The RNA-DNA hybrids thus formed in the wells were detected by an immunoenzymatic assay using a monoclonal antiRNA-DNA hybrid antibody. The sensitivity of this system was 2.7 x 10(4) cells ml(-1). This simple, sensitive and inexpensive hybridization and immunoenzymatic assay system should facilitate the detection of Campylobacter in food and clinical samples.
Lamoureux
M
M
Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada.
Fliss
L
L
Blais
B W
BW
Messier
S
S
Holley
R A
RA
Simard
R E
RE
D001431:Bacteriological Techniques; D016123:Campylobacter jejuni / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002876:Chromosomes, Bacterial / Q000235:genetics; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis*; D016678:Genome; D009693:Nucleic Acid Hybridization / Q000379:methods; D010969:Plastics; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004269:DNA, Bacterial; D010969:Plastics