open access

https://hdl.handle.net/2115/92973

Analysis of Unamplified 16S rRNA of Ammonia-Oxidizing Bacteria in Activated Sludge by Spectrophotometry Using Gold Nanoprobes

英語

heating

2023-09-08

American Chemical Society

ACS Es&t Water

3(9)

3113-3120

The identification of the bacteria contributing to the water purification processes in these systems is crucial for the development and optimization of wastewater treatment systems by engineers. We developed a simple assay for the colorimetric quantification of bacterial 16S rRNA of functional bacteria extracted from wastewater samples without reverse transcription of RNA or quantitative polymerase chain reaction (qPCR). The heat treatment of the test sample significantly reduced the reaction time (<3 min) and improved the assay sensitivity. Following assay optimization, a calibration curve for 16S rRNA of ammonia-oxidizing bacteria (AOB) of the beta subdivision of the class Proteobacteria was created using 16S rRNA extracted from activated sludge samples. The curve was linear above 1.9 x 10(6) copies/mL of 16S rRNA of AOB, which was 2 orders of magnitude lower than that in our previous studies. Finally, we determined the abundance of 16S rRNA of AOB in activated sludge samples using the calibration curve and compared the results obtained by reverse transcription-qPCR. There was some correlation in the RNA concentrations of AOB determined by both methods. Thus, we have successfully developed an innovative tool to quantify the 16S rRNA concentration of AOB in wastewater treatment processes.

学術雑誌論文

Accepted Manuscript

This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS ES&T Water, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/articlesonrequest/AOR-WYZ9M3GIUGZB4PY8N5AN.