Citation: HE Hao-Yan, HUANG En-Qi, LI Zhu-Jun, SHU Bo-Wen, XU Bang-Lao, LIU Da-Yu. Droplet Digital Microfluidic Assay for Rapid Bacterium Quantitation[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(7): 855-862. doi: 10.19756/j.issn.0253-3820.201022 shu

Droplet Digital Microfluidic Assay for Rapid Bacterium Quantitation

HE Hao-Yan ^1, ,
HUANG En-Qi ¹ ,
LI Zhu-Jun ¹ ,
SHU Bo-Wen ^1,2,3 ,
XU Bang-Lao ^1,2,3 ,
LIU Da-Yu ^1,2,3

1.
Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou 510180, China;
2.
Guangdong Engineering Technology Research Center of Microfluidic Chip Medical Diagnosis, Guangzhou 510180, China;
3.
Clinical Molecular Medicine and Molecular Diagnosis Key Laboratory of Guangdong Province, Guangzhou 510180, China

Received Date: 9 January 2020
Revised Date: 24 February 2020

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 81871726, 21804044).

Currently available bacterium quantitation assays majorly depend on specialized facilities and associate with long turnaround time. To overcome these limitations, a droplet digital microfluidic assay was developed for rapid bacterium quantitation. The microfluidic assay featured by syringe vacuum activated droplet generation, by which stochastic single cell confinement in monodisperse picoliter volumes in parallel modules was achieved. The presence of live bacteria in droplets was indicated by the change of fluorescence intensity due to the chromogenic reaction caused by live bacteria. Bacteria density in the original sample was calculated by fitting the proportion of positives (p) to the Poisson distribution algorithm. The total analytical process for bacteria quantitation was completed in 3.5 h. The dynamic detection range was 10⁵-10⁸ CFU/mL, with relative standard deviation <5%. With advantages of simple operation and short analysis time, the developed microfluidic assay was expected to be adapted for rapid bacterium quantitation.
- Microfluidic chip,
- Droplet digital assay,
- Syringe vacuum,
- Bacterium,
- Quantitation detection
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