期刊
ACS SYNTHETIC BIOLOGY
卷 10, 期 5, 页码 1132-1142出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.0c00640
关键词
whole-cell biosensor; bioluminescence; luxCDABE; blood cells; hematuria; single-photon avalanche photodiode
资金
- Israel Ministry of Science and Technology [3-14364]
- Israel Science Foundation [1158/17]
Early detection of blood in urine is crucial for the treatment of serious diseases, and the development of low-cost portable biosensors for this purpose is necessary. An ultrasensitive whole-cell bacterial biosensor was designed for heme detection in urine, showing promising results for hematuria detection.
The early detection of blood in urine (hematuria) can play a crucial role in the treatment of serious diseases (e.g., infections, kidney disease, schistosomiasis, and cancer). Therefore, the development of low-cost portable biosensors for blood detection in urine has become necessary. Here, we designed an ultrasensitive whole-cell bacterial biosensor interfaced with an optoelectronic measurement module for heme detection in urine. Heme is a red blood cells (RBCs) component that is liberated from lysed cells. The bacterial biosensor includes Escherichia coli cells carrying a heme-sensitive synthetic promoter integrated with a luciferase reporter (luxCDABE) from Photorhabdus luminescens. To improve the bacterial biosensor performance, we re-engineered the genetic structure of luxCDABE operon by splitting it into two parts (luxCDE and luxAB). The luxCDE genes were regulated by the heme-sensitive promoter, and the luxAB genes were regulated by either constitutive or inducible promoters. We examined the genetic circuit's performance in synthetic urine diluent supplied with heme and in human urine supplied with lysed blood. Finally, we interfaced the bacterial biosensor with a light detection setup based on a commercial optical measurement single-photon avalanche photodiode (SPAD). The whole-cell biosensor was tested in human urine with lysed blood, demonstrating a low-cost, portable, and easy-to-use hematuria detection with an ON-to-OFF ratio of 6.5-fold for blood levels from 5 x 10(4) to 5 x 10(5) RBC per mL of human urine.
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