Detection of Salmonella spp. in food products by polymerase chain reaction and hybridization assay in microplate format.
Here, hybridization assay of amplified products is described which detect Salmonella spp. from chicken fillets and other food homogenates within 24 h. This technique is composed of four steps: (1) sample is pre-enriched overnight in phosphate buffered peptone water; (2) total DNA is extracted; (3) a Salmonella spp. specific DNA target sequence is amplified by polymerase chain reaction; (4) amplified products are captured by a probe covalently bound onto NH-Covalink (Nunc, Danemark) microwells and detected by a chemiluminescent enzymatic reaction. This hybridization of amplified products was demonstrated as sensitive as their analysis on agarose gel. Compared to a bacteriological method for Salmonella spp. detection, its specificity was estimated at 100% and its sensitivity was 93.2% from analysis of 207 naturally contaminated chicken fillets samples.
Soumet
C
C
Unité de Recherches de Biologie Moléculaire, CNEVA Ploufragan, Zoopôle Les Croix, France.
Ermel
G
G
Salvat
G
G
Colin
P
P
D000818:Animals; D002645:Chickens; D005516:Food Microbiology; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012475:Salmonella / Q000302:isolation & purification*