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Current Topics in Analytical Chemistry   Volumes    Volume 15 
A chemiluminometric inorganic pyrophosphate ion assay using inorganic pyrophosphatase, pyruvate oxidase, and isoluminol for specific detection of E. coli RNA
Hideaki Nakamura, Noriko Suzuki, Sachie Okubo, Koji Karasawa, Masaru Kato, Hidetoshi Arakawa
Pages: 45 - 51
Number of pages: 7
Current Topics in Analytical Chemistry
Volume 15 

Copyright © 2023 Research Trends. All rights reserved

In the present study, we attempted to detect pathogenic microbes employing the analyte of inorganic pyrophosphate ion (PPi) generated by nucleic acid reaction. We employed a nucleic acid sequence-based amplification (NASBA) method for amplification of RNA (in 16S rRNA) extracted from E. coli which was used as a substitute for pathogenic microbes. PPi was detected as the product of the NASBA reaction by a chemiluminometric inorganic pyrophosphate ion (CL-PPi) assay which consists of three enzymatic reactions of inorganic pyrophosphatase (IP), pyruvate oxidase (PO), and isoluminol-microperoxidase chemiluminescence. Using our method, the difference between the negative control and positive control (E. coli RNA) was detected.
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