Anti-DNA.RNA antibodies: an efficient tool for non-isotopic detection of Listeria species through a liquid-phase hybridization assay.
This study was undertaken to evaluate the potential of a new approach using anti-DNA.RNA monoclonal antibodies to detect Listeria in both pure culture and inoculated meat and meat products. A sensitive liquid-phase assay was first developed, based on the formation in solution of a hybrid between a 784-bp DNA probe, specific for the genus Listeria, and target rRNA. Monoclonal antibody and antisera raised against hybrid nucleic acids were then used in various immunoenzymatic assays to detect specific hybrids formed in solution. System 2, using a double sandwich enzyme-linked immunosorbent assay, and system 1, using a biotinylated probe, proved to be very effective. The method using biotin-streptavidin complex, however, resulted in a higher background signal. System 2 described here, using unlabeled probe, was more effective. This strategy allowed the detection of as little as 2.5 pg target RNA from pure culture and 500 cells from inoculated meat homogenate, even in the presence of other contaminating bacteria. The assay was more sensitive and could be completed within 3 h, as opposed to several days when conventional culture methods were used.
Fliss
I
I
Département de Sciences et Technologie des aliments, Université Laval, Sainte-Foy, Québec, Canada.
St Laurent
M
M
Emond
E
E
Simard
R E
RE
Lemieux
R
R
Ettriki
A
A
Pandian
S
S
D000911:Antibodies, Monoclonal; D015342:DNA Probes; D007124:Immunoenzyme Techniques; D008087:Listeria / Q000302:isolation & purification*; D008460:Meat / Q000382:microbiology; D009693:Nucleic Acid Hybridization; D012335:RNA, Ribosomal / Q000235:genetics*; D012680:Sensitivity and Specificity; D013696:Temperature
D000911:Antibodies, Monoclonal; D015342:DNA Probes; D012335:RNA, Ribosomal