PMID- 36213081
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20221011
IS  - 2296-4185 (Print)
IS  - 2296-4185 (Electronic)
IS  - 2296-4185 (Linking)
VI  - 10
DP  - 2022
TI  - Microfluidics combined with fluorescence in situ hybridization (FISH) for Candida 
      spp. detection.
PG  - 987669
LID - 10.3389/fbioe.2022.987669 [doi]
LID - 987669
AB  - One of the most prevalent healthcare-associated infection is the urinary tract 
      infection (UTI), caused by opportunistic pathogens such as Candida albicans or 
      non-albicans Candida species (NACS). Urine culture methods are routinely used for 
      UTI diagnostics due to their specificity, sensitivity and low-cost. However, 
      these methods are also laborious, time- and reagent-consuming. Therefore, 
      diagnostic methods relying on nucleic acids have been suggested as alternatives. 
      Nucleic acid-based methods can provide results within 24 h and can be adapted to 
      point-of-care (POC) detection. Here, we propose to combine fluorescence in situ 
      hybridization (FISH) with a microfluidic platform for the detection of Candida 
      spp. As a case study we used C. tropicalis, which is reported as the second most 
      common NACS urine isolate obtained from patients suspected with UTI. The 
      microfluidic platform proposed in this study relies on hydrodynamic trapping, and 
      uses physical barriers (e.g., microposts) for the separation of target cells from 
      the suspension. Using a specific peptide nucleic acid (PNA) probe, the FISH 
      procedure was applied onto previously trapped C. tropicalis cells present inside 
      the microfluidic platform. Fluorescence signal intensity of hybridized cells was 
      captured directly under the epifluorescence microscope. Overall, the PNA probe 
      successfully detected C. tropicalis in pure culture and artificial urine (AU) 
      using FISH combined with the microfluidic platform. Our findings reveal that FISH 
      using nucleic acid mimics (PNA) in combination with microfluidics is a reliable 
      method for the detection of microorganisms such as C. tropicalis. As such, this 
      work provides the basis for the development of a POC detection platform in the 
      future.
CI  - Copyright (c) 2022 Barbosa, Rodrigues, Cerqueira, Miranda and Azevedo.
FAU - Barbosa, Violina Baranauskaite
AU  - Barbosa VB
AD  - LEPABE-Laboratory for Process Engineering, Environment, Biotechnology and Energy, 
      Department of Chemical Engineering, Faculty of Engineering of University of 
      Porto, Porto, Portugal.
AD  - ALiCE-Associate Laboratory in Chemical Engineering, Faculty of Engineering, 
      University of Porto, Porto, Portugal.
FAU - Rodrigues, Celia F
AU  - Rodrigues CF
AD  - LEPABE-Laboratory for Process Engineering, Environment, Biotechnology and Energy, 
      Department of Chemical Engineering, Faculty of Engineering of University of 
      Porto, Porto, Portugal.
AD  - ALiCE-Associate Laboratory in Chemical Engineering, Faculty of Engineering, 
      University of Porto, Porto, Portugal.
FAU - Cerqueira, Laura
AU  - Cerqueira L
AD  - LEPABE-Laboratory for Process Engineering, Environment, Biotechnology and Energy, 
      Department of Chemical Engineering, Faculty of Engineering of University of 
      Porto, Porto, Portugal.
AD  - ALiCE-Associate Laboratory in Chemical Engineering, Faculty of Engineering, 
      University of Porto, Porto, Portugal.
FAU - Miranda, Joao M
AU  - Miranda JM
AD  - ALiCE-Associate Laboratory in Chemical Engineering, Faculty of Engineering, 
      University of Porto, Porto, Portugal.
AD  - CEFT-Transport Phenomena Research Center, Department of Chemical Engineering, 
      Faculty of Engineering of University of Porto, Porto, Portugal.
FAU - Azevedo, Nuno F
AU  - Azevedo NF
AD  - LEPABE-Laboratory for Process Engineering, Environment, Biotechnology and Energy, 
      Department of Chemical Engineering, Faculty of Engineering of University of 
      Porto, Porto, Portugal.
AD  - ALiCE-Associate Laboratory in Chemical Engineering, Faculty of Engineering, 
      University of Porto, Porto, Portugal.
LA  - eng
PT  - Journal Article
DEP - 20220923
PL  - Switzerland
TA  - Front Bioeng Biotechnol
JT  - Frontiers in bioengineering and biotechnology
JID - 101632513
PMC - PMC9539416
OTO - NOTNLM
OT  - C. tropicalis
OT  - FISH
OT  - UTI
OT  - detection
OT  - microfluidics
COIS- The authors declare that the research was conducted in the absence of any 
      commercial or financial relationships that could be construed as a potential 
      conflict of interest.
EDAT- 2022/10/11 06:00
MHDA- 2022/10/11 06:01
PMCR- 2022/01/01
CRDT- 2022/10/10 04:24
PHST- 2022/07/06 00:00 [received]
PHST- 2022/08/08 00:00 [accepted]
PHST- 2022/10/10 04:24 [entrez]
PHST- 2022/10/11 06:00 [pubmed]
PHST- 2022/10/11 06:01 [medline]
PHST- 2022/01/01 00:00 [pmc-release]
AID - 987669 [pii]
AID - 10.3389/fbioe.2022.987669 [doi]
PST - epublish
SO  - Front Bioeng Biotechnol. 2022 Sep 23;10:987669. doi: 10.3389/fbioe.2022.987669. 
      eCollection 2022.