An integrated microfluidic system for early detection of sepsis-inducing bacteria

被引:42
作者
Fang, Yen-Ling [1 ]
Wang, Chih-Hung [1 ]
Chen, Yi-Sin [1 ]
Chien, Chun-Chih [2 ]
Kuo, Feng-Chih [3 ]
You, Huey-Ling [2 ]
Lee, Mel S. [3 ]
Lee, Gwo-Bin [1 ,4 ,5 ]
机构
[1] Natl Tsing Hua Univ, Dept Power Mech Engn, Hsinchu 30013, Taiwan
[2] Kaohsiung Chang Gung Mem Hosp, Lab Med, Kaohsiung, Taiwan
[3] Chang Gung Univ, Kaohsiung Chang Gung Mem Hosp, Dept Orthopaed Surg, Kaohsiung 83301, Taiwan
[4] Natl Tsing Hua Univ, Inst Biomed Engn, Hsinchu, Taiwan
[5] Natl Tsing Hua Univ, Inst Nano Engn & Microsyst, Hsinchu, Taiwan
关键词
POLYMERASE-CHAIN-REACTION; RED-BLOOD-CELLS; AMPLIFICATION; CHIP; DIAGNOSIS; MEMBRANE; SIZE;
D O I
10.1039/d0lc00966k
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Since early diagnosis of sepsis may assist clinicians in initiating timely, effective, and prognosis-improving antibiotic therapy, we developed an integrated microfluidic chip (IMC) for rapid isolation of both Gram-positive and Gram-negative bacteria from blood. The device comprised a membrane-based filtration module (90 min operating time), a bacteria-capturing module using a micro-mixer containing magnetic beads coated with "flexible neck" regions of mannose-binding lectin proteins for bacteria capture (20 min), and a miniature polymerase chain reaction (PCR) module for bacteria identification (90 min via TaqMan (R) probe technology). The filter separated all white blood cells and 99.5% of red blood cells from bacteria, which were captured at rates approaching 85%. The PCR assay's limit of detection was 5 colony-forming units (CFU) per reaction, and the entire process was completed in only 4 h. Since this is far less than that for culture-based approaches, this IMC may serve as a promising device for detection of sepsis.
引用
收藏
页码:113 / 121
页数:9
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