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A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay

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Abstract

A high-throughput microfluidic poly-(dimethylsiloxane) biochip was developed to quantify bacterial adhesion to single host cells by real-time PCR assay. The biochip is simply structured with a two-dimensional array of 900 micro-wells, one inlet, and one outlet micro-channels. Isolation of single infected host cells into the individual micro-wells of the biochip was achieved by one-step vacuum-driven microfluidics. The adhered bacterial cells were then quantified by direct on-chip real-time PCR assay with single-bacterium-detection sensitivity. The performance of this microfluidic platform was demonstrated through profiling of the association of a common bacterial pathogen, Pseudomonas aeruginosa, to single host human lung epithelial A549 cells, revealing an adherence distribution that has not been previously reported. This microfluidic platform offers a simple and effective tool for biologists to analyze pathogen–host interaction at the single-cell level without the necessities of fluorescence labeling. The chip can similarly be used for other PCR-based applications requiring single-cell analysis.

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Acknowledgments

The authors thank Star Array Pte Ltd, Singapore, for the technical support.

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Correspondence to Hai-Qing Gong.

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Zhang, R., Gong, HQ., Zeng, X.D. et al. A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay. Anal Bioanal Chem 405, 4277–4282 (2013). https://doi.org/10.1007/s00216-013-6826-0

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