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PMID: 12902306 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Detection of bacteria carrying the stx2 gene by in situ loop-mediated isothermal amplification.

Applied and environmental microbiology ·Vol. 69 ·No. 8 ·2003-08-00 ·Pages 5023-8

Maruyama F, Kenzaka T, Yamaguchi N, Tani K, Nasu M

Abstract

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.

MeSH Terms
Escherichia coli O157/genetics Gene Amplification In Situ Hybridization, Fluorescence Polymerase Chain Reaction Shiga Toxin 2/genetics
Chemicals
Shiga Toxin 2
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Maruyama Fumito
Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Kenzaka Takehiko
Yamaguchi Nobuyasu
Tani Katsuji
Nasu Masao
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.69.8.5023-5028.2003
Published
2003-08-00
Pages
5023-8
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC169117
Subset
IM
External Links
PubMed source DOI: 10.1128/AEM.69.8.5023-5028.2003 PMC Full Text View journal details
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