A PCR-microplate capture hybridization method to detect Listeria monocytogenes in blood.
In order to improve the diagnosis of Listeria monocytogenes infection, we have developed a polymerase chain reaction (PCR)-based assay combined with microplate capture hybridization technique. The system is based on selective amplification of L. monocytogenes with two specific primers based on the iap gene. The amplicon produced, with digoxigenin 11-dUTP incorporated during PCR, is hybridized in streptavidin-coated microtitre plates prepared with biotinylated specific DNA probe. The method involved requires approximately 6-8 h, and its high sensitivity, rapidity and simplicity should make it valuable for diagnosis and for epidemiological studies of listeriosis.
Cocolin
L
L
Dipartimento di Scienze degli Alimenti, Facoltà di Agraria, Università di Udine, Italy.
Manzano
M
M
Cantoni
C
C
Comi
G
G
D000818:Animals; D016470:Bacteremia / Q000175:diagnosis / Q000382:microbiology; D001769:Blood / Q000382:microbiology*; D015342:DNA Probes; D004076:Digoxigenin; D006801:Humans; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods*; D008089:Listeria monocytogenes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004076:Digoxigenin