Chemiluminescent and colorimetric enzymatic assays for the detection of PCR-amplified Salmonella sp. products in microplates.
To improve Salmonella detection, we developed nonradioactive hybridization assays of amplified products from pure Salmonella cultures. Biotin-labeled PCR products were trapped by internal probes covalently bound to CovaLink-NH MicroWells and detected by colorimetric or chemiluminescent enzymatic reactions. The sensitivities of colorimetric assays using peroxidase and alkaline phosphatase were similar to those obtained with an ethidium bromide-stained agarose gel; both procedures allow the detection of 50 Salmonella cells. Chemiluminescence was 10-fold more sensitive than colorimetry.
Soumet
C
C
Centre National d'Etudes Veterinaires et Alimentaires Ploufragan, France.
Ermel
G
G
Boutin
P
P
Boscher
E
E
Colin
P
P
D000469:Alkaline Phosphatase; D003124:Colorimetry / Q000706:statistics & numerical data; D004396:Coloring Agents; D004269:DNA, Bacterial / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D004996:Ethidium; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization; D009195:Peroxidase; D016133:Polymerase Chain Reaction; D012475:Salmonella / Q000235:genetics*; D012680:Sensitivity and Specificity
D004396:Coloring Agents; D004269:DNA, Bacterial; D009195:Peroxidase; D000469:Alkaline Phosphatase; D004996:Ethidium