Abstract
The prerequisites for rapid screening of total bacteria in drinking water are low detection limit and convenience. Inspired by commercial adenosine 5′-triphosphate (ATP) based total bacterial detection kits, we pursued likewise convenience but with much lower detection limit. Existing intercalation fluorescence-based techniques employ multiple reagents to permeate the cell membrane and intercalate dye into the DNA in discrete sequential steps. A simple multi-functional reagent is proposed to do the same within one step. Surfactants (TritonX and SDS), and intercalating dyes (SYBR green, SYBR gold) were examined for their mutual compatibility and augmented with EDTA. Evaluation was performed with Gram negative Escherichia coli K12 (E. coli K12) and Gram positive Bacillus subtilis (B. subtilis) at serial dilution ratios from 10−6 to 10−2. Comparison was made with absorbance (600 nm) measurements and a commercial ATP kit. Using charge integrated photodetection, the proposed 1-step reagent achieved an LOD (1.00 × 10−6, B. subtilis) that is two orders of magnitude lower than that of ATP kit (LOD = 1.06× 10−4). This means it could detect minute quantity of total bacteria that is otherwise undetected by the ATP kit.
Original language | English |
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Article number | 137541 |
Journal | Chemosphere |
Volume | 313 |
DOIs | |
State | Published - Feb 2023 |
Bibliographical note
Funding Information:This study was supported by the National Research Foundation of Korea ( NRF-2019R1A2C2084233 and NRF-2022R1F1A1063936 ).
Publisher Copyright:
© 2022 Elsevier Ltd
Keywords
- Charge integrated photodetection
- Convenience
- Intercalated dye fluorescence
- Low detection limit
- Multifunctional reagent
- Total bacteria in drinking water