PMID- 22212757
OWN - NLM
STAT- MEDLINE
DCOM- 20120814
LR  - 20120131
IS  - 1872-8359 (Electronic)
IS  - 0167-7012 (Linking)
VI  - 88
IP  - 2
DP  - 2012 Feb
TI  - Capture antibody targeted fluorescence in situ hybridization (CAT-FISH): dual 
      labeling allows for increased specificity in complex samples.
PG  - 275-84
LID - 10.1016/j.mimet.2011.12.009 [doi]
AB  - Pathogen detection using biosensors is commonly limited due to the need for 
      sensitivity and specificity in detecting targets within mixed populations. These 
      issues were addressed through development of a dual labeling method that allows 
      for both liquid-phase fluorescence in situ hybridization (FISH) and capture 
      antibody targeted detection (CAT-FISH). CAT-FISH was developed using Escherichia 
      coli O157:H7 and Staphylococcus aureus as representative bacteria, and processing 
      techniques were evaluated with regard to FISH intensities and antibody 
      recognition. The alternative fixative solution, methacarn, proved to be superior 
      to standard solid-phase paraformaldehyde fixation procedures, allowing both FISH 
      labeling and antibody recognition. CAT-FISH treated cells were successfully 
      labeled with FISH probes, captured by immunomagnetic separation using fluorescent 
      cytometric array beads, and detected using a cytometric array biosensor. CAT-FISH 
      treated cells were detectable with LODs comparable to the standard antibody-based 
      technique, (~10(3)cells/ml in PBS), and the technique was also successfully 
      applied to two complex matrices. Although immunomagnetic capture and detection 
      using cytometric arrays were demonstrated, CAT-FISH is readily applicable to any 
      antibody-based fluorescence detection platform, and further optimization for 
      sensitivity is possible via inclusion of fluorescently tagged antibodies. Since 
      the confidence level needed for positive identification of a detected target is 
      often paramount, CAT-FISH was developed to allow two separate levels of 
      specificity, namely nucleic acid and protein signatures. With proper selection of 
      FISH probes and capture antibodies, CAT-FISH may be used to provide rapid 
      detection of target pathogens from within complex matrices with high levels of 
      confidence.
CI  - Copyright (c) 2011 Elsevier B.V. All rights reserved.
FAU - Stroot, Joyce M
AU  - Stroot JM
AD  - Department of Cell Biology, Microbiology, and Molecular Biology, University of 
      South Florida, 4202 E. Fowler Ave. ISA2015, Tampa, FL 33620, USA. 
      jstroot@cirisenergy.com
FAU - Leach, Kelly M
AU  - Leach KM
FAU - Stroot, Peter G
AU  - Stroot PG
FAU - Lim, Daniel V
AU  - Lim DV
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20111222
PL  - Netherlands
TA  - J Microbiol Methods
JT  - Journal of microbiological methods
JID - 8306883
RN  - 0 (Antibodies, Bacterial)
RN  - 0 (Fixatives)
SB  - IM
MH  - Animals
MH  - Antibodies, Bacterial/metabolism
MH  - Bacteriological Techniques/*methods
MH  - Biosensing Techniques
MH  - Blood/microbiology
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - Escherichia coli O157/chemistry/isolation & purification
MH  - Fixatives
MH  - Fluorescent Antibody Technique/methods
MH  - Immunomagnetic Separation/*methods
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Protein Binding
MH  - Sensitivity and Specificity
MH  - Sheep
MH  - Spinacia oleracea/microbiology
MH  - Staphylococcus aureus/chemistry/isolation & purification
EDAT- 2012/01/04 06:00
MHDA- 2012/08/15 06:00
CRDT- 2012/01/04 06:00
PHST- 2011/11/03 00:00 [received]
PHST- 2011/12/13 00:00 [revised]
PHST- 2011/12/13 00:00 [accepted]
PHST- 2012/01/04 06:00 [entrez]
PHST- 2012/01/04 06:00 [pubmed]
PHST- 2012/08/15 06:00 [medline]
AID - S0167-7012(11)00435-0 [pii]
AID - 10.1016/j.mimet.2011.12.009 [doi]
PST - ppublish
SO  - J Microbiol Methods. 2012 Feb;88(2):275-84. doi: 10.1016/j.mimet.2011.12.009. 
      Epub 2011 Dec 22.