Detection of bacteria carrying the stx2 gene by in situ loop-mediated isothermal amplification.
A new in situ DNA amplification technique for microscopic detection of bacteria carrying a specific gene is described. Loop-mediated isothermal amplification (LAMP) was used to detect stxA(2) in Escherichia coli O157:H7 cells. The mild permeabilization conditions and low isothermal temperature used in the in situ LAMP method caused less cell damage than in situ PCR. It allowed use of fluorescent antibody labeling in the bacterial mixture after the DNA amplification for identification of E. coli O157:H7 cells with an stxA(2) gene. Higher-contrast images were obtained with this method than with in situ PCR.
Maruyama
Fumito
F
Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Kenzaka
Takehiko
T
Yamaguchi
Nobuyasu
N
Tani
Katsuji
K
Nasu
Masao
M
D019453:Escherichia coli O157 / Q000235:genetics*; D005784:Gene Amplification; D017404:In Situ Hybridization, Fluorescence; D016133:Polymerase Chain Reaction; D022641:Shiga Toxin 2 / Q000235:genetics*
D022641:Shiga Toxin 2