PMID- 32003380
OWN - NLM
STAT- MEDLINE
DCOM- 20210621
LR  - 20210621
IS  - 1473-0189 (Electronic)
IS  - 1473-0189 (Linking)
VI  - 20
IP  - 5
DP  - 2020 Mar 3
TI  - A "sample-in-multiplex-digital-answer-out" chip for fast detection of pathogens.
PG  - 979-986
LID - 10.1039/c9lc01143a [doi]
AB  - Point-of-care (POC) testing offers rapid diagnostic results. However, the 
      quantification of current methods is performed using standard curves and external 
      references, and not direct and absolute quantification. This paper describes an 
      integrated multiplex digital recombinase polymerase amplification (ImdRPA) 
      microfluidic chip which combines DNA extraction, multiplex digital RPA and 
      fluorescence detection together in one chip, creating a 
      "sample-in-multiplex-digital-answer-out" system. Multi-layer soft lithography 
      technology was used, with polydimethylsiloxane (PDMS) as the chip material and a 
      glass slide as the substrate. This microfluidic chip has a six-layer structure 
      and screw microvalve control function. The sample preparation for the chip 
      involved magnetic bead-based nucleic acid extraction, which was completed within 
      15 min without any instrument dependence. The dRPA region was divided into 4 
      regions (3 positive detection areas and 1 negative control area) and included a 
      total of 12 800 chambers, with each chamber being able to contain a volume of 2.7 
      nL. The screw valve allowed for the reaction components of each specific goal to 
      be pre-embedded in different regions of the chambers. The reagents were passively 
      driven into the dRPA region using vacuum-based self-priming introduction. 
      Furthermore, we successfully demonstrated that the chip can simultaneously detect 
      three species of pathogenic bacteria within 45 min and give digital quantitative 
      results without the need to establish a standard curve in contaminated milk. 
      Moreover, the detection limit of this ImdRPA microfluidic chip was found to be 10 
      bacterial cells for each kind of pathogen. These characteristics enhance its 
      applicability for rapid detection of foodborne bacteria at the point-of-care 
      (POC). We envision that the further development of this integrated chip will lead 
      to rapid, multiplex and accurate detection of foodborne bacteria in a feasible 
      manner.
FAU - Yin, Juxin
AU  - Yin J
AUID- ORCID: 0000-0002-4271-5157
AD  - Research Centre for Analytical Instrumentation, Institute of Cyber-Systems and 
      Control, State Key Laboratory of Industrial Control Technology, Zhejiang 
      University, Hangzhou, Zhejiang Province 310058, P. R. China. muying@zju.edu.cn 
      and Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, 
      National Ministry of Education), The Second Affiliated Hospital, School of 
      Medicine, Zhejiang University, Hangzhou 310009, China and Institute of 
      Translational Medicine, Zhejiang University, Hangzhou 310029, China.
FAU - Zou, Zheyu
AU  - Zou Z
AD  - Research Centre for Analytical Instrumentation, Institute of Cyber-Systems and 
      Control, State Key Laboratory of Industrial Control Technology, Zhejiang 
      University, Hangzhou, Zhejiang Province 310058, P. R. China. muying@zju.edu.cn 
      and College of Life Sciences, Zhejiang University, Hangzhou 310058, Zhejiang, P. 
      R. China.
FAU - Hu, Zhenming
AU  - Hu Z
AD  - Research Centre for Analytical Instrumentation, Institute of Cyber-Systems and 
      Control, State Key Laboratory of Industrial Control Technology, Zhejiang 
      University, Hangzhou, Zhejiang Province 310058, P. R. China. muying@zju.edu.cn.
FAU - Zhang, Shan
AU  - Zhang S
AD  - Research Centre for Analytical Instrumentation, Institute of Cyber-Systems and 
      Control, State Key Laboratory of Industrial Control Technology, Zhejiang 
      University, Hangzhou, Zhejiang Province 310058, P. R. China. muying@zju.edu.cn 
      and College of Life Sciences, Zhejiang University, Hangzhou 310058, Zhejiang, P. 
      R. China.
FAU - Zhang, Fengping
AU  - Zhang F
AD  - HeZe Municipal hospital, HeZe 274000, China.
FAU - Wang, Ben
AU  - Wang B
AUID- ORCID: 0000-0002-4134-1835
AD  - Cancer Institute (Key Laboratory of Cancer Prevention and Intervention, National 
      Ministry of Education), The Second Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310009, China and Institute of Translational 
      Medicine, Zhejiang University, Hangzhou 310029, China.
FAU - Lv, Shaowu
AU  - Lv S
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, College of Life Science, Jilin University, Changchun 130000, China. 
      lvsw@jlu.edu.cn.
FAU - Mu, Ying
AU  - Mu Y
AUID- ORCID: 0000-0002-4322-2522
AD  - Research Centre for Analytical Instrumentation, Institute of Cyber-Systems and 
      Control, State Key Laboratory of Industrial Control Technology, Zhejiang 
      University, Hangzhou, Zhejiang Province 310058, P. R. China. muying@zju.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Lab Chip
JT  - Lab on a chip
JID - 101128948
RN  - 0 (Recombinases)
SB  - IM
MH  - Lab-On-A-Chip Devices
MH  - *Microfluidics
MH  - Nucleic Acid Amplification Techniques
MH  - Oligonucleotide Array Sequence Analysis
MH  - Point-of-Care Systems
MH  - Point-of-Care Testing
MH  - *Recombinases
EDAT- 2020/02/01 06:00
MHDA- 2021/06/22 06:00
CRDT- 2020/02/01 06:00
PHST- 2020/02/01 06:00 [pubmed]
PHST- 2021/06/22 06:00 [medline]
PHST- 2020/02/01 06:00 [entrez]
AID - 10.1039/c9lc01143a [doi]
PST - ppublish
SO  - Lab Chip. 2020 Mar 3;20(5):979-986. doi: 10.1039/c9lc01143a.