RRC ID 50856
著者 Haruta S, Iino T, Ohkuma M, Suzuki KI, Igarashi Y.
タイトル Ca2+ in Hybridization Solutions for Fluorescence in situ Hybridization Facilitates the Detection of Enterobacteriaceae.
ジャーナル Microbes Environ
Abstract Fluorescence in situ hybridization (FISH) has been employed to identify microorganisms at the single cell level under a microscope. Extensive efforts have been made to improve and extend the FISH technique; however, the development of a widely applicable protocol is a continuing challenge. The present study evaluated the effects of divalent cations in the hybridization solution on the FISH-based detection of various species of bacteria and archaea with rRNA-targeted probes. A flow cytometric analysis after FISH with a standard hybridization buffer detected positive signals from less than 30% of Escherichia coli IAM 1264 cells. However, the number of cells with positive signals increased to more than 90% after the addition of calcium chloride to the hybridization buffer. Mn2+ also had positive effects, whereas Mg2+ did not. The positive effects of Ca2+ were similarly observed for bacteria belonging to Enterobacteriaceae, including Enterobacter sakazakii IAM 12660T, E. aerogenes IAM 12348, Klebsiella planticola IAM 14202, and Salmonella enterica subsp. enterica serovar Typhimurium strain LT2. These results indicate that the supplementation of Ca2+ to the hybridization buffer for FISH contributes to the efficient detection of Enterobacteriaceae cells.
巻・号 32(2)
ページ 142-146
公開日 2017-6-24
DOI 10.1264/jsme2.ME16186
PMID 28515389
PMC PMC5478537
MeSH Calcium / chemistry* Enterobacteriaceae / isolation & purification* In Situ Hybridization, Fluorescence* Oligonucleotide Probes
IF 2.575
引用数 2
リソース情報
一般微生物 JCM 1668 JCM 14847 JCM 1149