Detection of Bacteria Carrying the stx 2 Gene by In Situ Loop-Mediated Isothermal Amplification-Reference-Cited by-同舟云学术

Detection of Bacteria Carrying the stx 2 Gene by In Situ Loop-Mediated Isothermal Amplification

Published:2003-08 Issue:8 Volume:69 Page:5023-5028
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Maruyama Fumito¹,Kenzaka Takehiko¹,Yamaguchi Nobuyasu¹,Tani Katsuji¹,Nasu Masao¹

Affiliation:

1. Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan

Abstract

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.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/AEM.69.8.5023-5028.2003

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