Development, validation, and standardization of polymerase chain reaction-based detection of E. coli O157.
A diagnostic polymerase chain reaction assay was developed for the detection of E. coli O157 as the first part of a multicenter validation and standardization project. The assay is based on amplification of sequences of the rfbE O157 gene and includes an internal amplification control. The selectivity of the assay was evaluated against 155 strains, including 32 E. coli O157, 38 E. coli non-O157, and 85 non-E. coli. It was shown to be highly inclusive (100%) and exclusive (100%). The assay has a 100% detection probability of approximately 2 x 10(3) cells per reaction.
Justus-Liebig-University, Institute of Veterinary Food Sciences, Frankfurter str 92, 35392 Giessen, Germany.
Amir
A
Abdulmawjood
Michael
M
Bülte
Stefanie
S
Roth
Hahn
H
Schönenbrücher
Nigel
N
Cook
Annet E
AE
Heuvelink
Jeffrey
J
Hoorfar
D002238:Carbohydrate Epimerases / Q000235:genetics; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry; D019453:Escherichia coli O157 / Q000737:chemistry* / Q000235:genetics; D005062:European Union; D005504:Food Analysis / Q000592:standards*; D005798:Genes, Bacterial / Q000235:genetics; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D000637:Transaminases / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial; D000637:Transaminases; D002238:Carbohydrate Epimerases; C061272:perosamine synthetase