PMID- 33381652
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20210101
IS  - 2396-8923 (Electronic)
IS  - 2396-8923 (Linking)
VI  - 5
IP  - 1
DP  - 2020
TI  - Development of a fixation-free fluorescence in situ hybridization for the 
      detection of Salmonella species.
PG  - bpaa024
LID - 10.1093/biomethods/bpaa024 [doi]
LID - bpaa024
AB  - Salmonella is one of the most important infectious bacteria causing severe 
      gastroenteritis and deaths in humans and animals, and the prompt diagnosis is 
      crucial for effective control and treatment. The detection of Salmonella still 
      depends principally on culture-based methods, which is time-consuming and 
      laborious. Recently, polyhexamethylene biguanide (PHMB) was discovered to have 
      cellular delivery properties and its combination with the fluorescence in situ 
      hybridization (FISH) method was exploited for oligomer delivery and the rapid 
      detection of Salmonella spps in this study. Cell-associated fluorescence was 
      quantified using Volocity((R)) 3-D image analysis software (Volocity 6.3, 
      PerkinElmer, Inc.). PHMB complexed with fluorophore-labelled species-specific 
      oligomers permeabilized freshly grown viable strains of Salmonella cells and 
      mediated strong cell-associated fluorescence signals. This strategy further 
      enabled a fixation-free protocol and hybridization in a single reaction. Using 
      the modified FISH method, monoculture Salmonella strains were validated as well 
      as detected in artificially contaminated water and milk within a turnaround 
      period of 5 h. The method was observed to be comparable with the standard FISH 
      technique (sFISH; P > 0.05). The findings suggest that fixation-free delivery and 
      hybridization of oligomers using PHMB can provide a simplified and prompt 
      strategy for Salmonella detection at the species level, and promote early 
      management responses to the disease and appropriate antimicrobial therapy.
CI  - (c) The Author(s) 2020. Published by Oxford University Press.
FAU - Adebowale, Oluwawemimo
AU  - Adebowale O
AD  - Department of Pathobiology and Population Sciences, Royal Veterinary College, 
      London NW1 0TU, UK.
FAU - Good, Liam
AU  - Good L
AD  - Department of Pathobiology and Population Sciences, Royal Veterinary College, 
      London NW1 0TU, UK.
LA  - eng
PT  - Journal Article
DEP - 20201216
PL  - England
TA  - Biol Methods Protoc
JT  - Biology methods & protocols
JID - 101693064
PMC - PMC7756007
OTO - NOTNLM
OT  - Salmonella
OT  - bacterial diagnostics
OT  - fluorescence in situ hybridization
OT  - polyhexamethylene biguanide
EDAT- 2021/01/01 06:00
MHDA- 2021/01/01 06:01
PMCR- 2020/12/16
CRDT- 2020/12/31 05:27
PHST- 2020/08/31 00:00 [received]
PHST- 2020/10/28 00:00 [revised]
PHST- 2020/11/03 00:00 [accepted]
PHST- 2020/12/31 05:27 [entrez]
PHST- 2021/01/01 06:00 [pubmed]
PHST- 2021/01/01 06:01 [medline]
PHST- 2020/12/16 00:00 [pmc-release]
AID - bpaa024 [pii]
AID - 10.1093/biomethods/bpaa024 [doi]
PST - epublish
SO  - Biol Methods Protoc. 2020 Dec 16;5(1):bpaa024. doi: 10.1093/biomethods/bpaa024. 
      eCollection 2020.