Establishment and application of a novel fluorescence-based analytical method for the rapid detection of viable bacteria in different samples

Yin, Qiuyue; Nie, Minghua; Diwu, Zhenjun; Zhang, Yuting; Wang, Lei; Yin, Dan-Dan; Li, Lian-Cheng

doi:10.1039/d0ay01247e

Cited by 9 publications

(2 citation statements)

References 38 publications

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“…Most of the existing portable fluorometers [30][31][32][33][34] have a very higher limit of detection (7 to 10 5 CFU/mL) requiring complicated procedures i.e., preparation of magnetic nanoparticles and detection time is usually higher (around 30 minutes). On the other hand, our multichannel fluorometer has a LOD of <1 CFU/mL with incubation and filtration and detection can be done within as low as 20 seconds.…”

Section: Introductionmentioning

confidence: 99%

“…Most of the existing portable fluorometers − have a very higher LOD (7–10 5 cfu/mL) requiring complicated procedures, that is, preparation of magnetic nanoparticles, and detection time is usually higher (around 30 min). On the other hand, our multichannel fluorometer has an LOD of <1 cfu/mL with incubation and filtration and detection being done within as low as 20 s. This multichannel device has the ability for parallel testing of samples, which presents a significant improvement over our earlier prototype. , It can feature an inherent negative control and has the potential to detect anaerobes.…”

Section: Introductionmentioning

confidence: 99%

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Rapid Ultrasensitive and High-Throughput Bioburden Detection: Microfluidics and Instrumentation

et al. 2022

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Contamination detection often requires lengthy culturing steps to detect low-level bioburden. To increase the rate of detection and decrease the limit of detection (LOD), a system featuring microfluidics and a multichannel fluorometer has been developed. The 8-channel fluorometer enables parallel testing of multiple samples with LOD as low as < 1 CFU/mL. This low-cost system utilizes the slope of fluorescence intensity that serves as the criterion for bioburden detection. The redox indicator dye resazurin is used to monitor the presence of viable cells, in this study and is reduced to resorufin with a high quantum yield at 585 nm. The sample under investigation is spiked with resazurin, loaded in a special-design microfluidic cassette and the rate of change is observed via the fluorometer. The method was validated using primary E. coli culture in comparison with a spectrophotometer which served as the gold standard. An optimized assay based on LB media was developed. The impact on the assay sensitivity based on incubation and filtration steps was also explored. The assay is shown to pick up inadvertent contamination from test tubes and pipette tips showing its applicability in real-world settings. The data analysis demonstrated a comparable performance of the multichannel fluorometer vis-a-vis the conventional plate reader. The multichannel system is shown to detect bioburden presence in as low as 20 seconds for bacterial concentrations ≥ 5 CFU/mL after 6 hours of incubation. Considering its portability, low cost, simplicity of operation and relevant assay sensitivity, the system is well-positioned to detect low-level bioburden in the laboratory, pharmaceutical and field settings.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Rapid Ultrasensitive and High-Throughput Bioburden Detection: Microfluidics and Instrumentation

et al. 2022

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Illuminating Bacterial Contamination in Water Sources: The Power of Fluorescence-Based Methods

Modi,

Bhatt

et al. 2023

J Fluoresc

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Advances in detection methods for viable Salmonella spp.: current applications and challenges

Zhuang,

Gong,

Shen

et al. 2023

ANAL. SCI.

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Establishment and application of a novel fluorescence-based analytical method for the rapid detection of viable bacteria in different samples

Abstract: The present study described a rapid method for readily detecting the total numbers of viable bacteria cells in various samples (including surface water, solid inoculants and soil samples) using an...

Cited by 9 publications

References 38 publications

Rapid Ultrasensitive and High-Throughput Bioburden Detection: Microfluidics and Instrumentation

Rapid Ultrasensitive and High-Throughput Bioburden Detection: Microfluidics and Instrumentation

Illuminating Bacterial Contamination in Water Sources: The Power of Fluorescence-Based Methods

Advances in detection methods for viable Salmonella spp.: current applications and challenges

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