[Direct in situ PCR method for the detection of verotoxin-producing Escherichia coli].
Rapid detection of verotoxin-producing Escherichia coli at a single-cell level under an epifluorescence microscope without culturing processes was accomplished by using the direct in situ PCR technique. We used a DNA primer set for amplification of the slt-I and slt-II genes encoding respectively verotoxin 1 and 2 (EVT and EVS primers). The bacterial cells were detected specifically by the HNPP (2-hydroxy-3-naphthoic acid-2'-phenylanilide phosphate)/Fast Red TR reaction technique. The direct in situ PCR with HNPP/Fast Red TR technique is applicable to the detection of verotoxin-producing bacteria with the slt-I or slt-II gene in not only Escherichia coli O157 but also VTEC of other serotypes.
Kurokawa
K
K
Faculty of Pharmaceutical Sciences, Osaka University, Japan.
Tani
K
K
Nasu
M
M
D001427:Bacterial Toxins / Q000096:biosynthesis / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D019453:Escherichia coli O157 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction / Q000379:methods*; D022622:Shiga Toxin 1
D001427:Bacterial Toxins; D004269:DNA, Bacterial; D022622:Shiga Toxin 1