PMID- 15735784
OWN - NLM
STAT- MEDLINE
DCOM- 20050801
LR  - 20131121
IS  - 1464-0325 (Print)
IS  - 1464-0325 (Linking)
VI  - 7
IP  - 3
DP  - 2005 Mar
TI  - Real-time quantitative PCR with gene probe, fluorochrome and flow cytometry for 
      microorganism analysis.
PG  - 257-62
AB  - Microorganism concentrations and viability can be better understood and clarified 
      by using both culture and non-culture methods. Here, using pure suspensions of E. 
      coli, three non-culture methods, namely, flow cytometry (FCM), epifluorescence 
      microscopy (EFM), and real-time quantitative polymerase chain reaction (real-time 
      qPCR), were compared with a traditional culture-based method. Using 
      fluorocome-labeling methods with FCM and EFM applications, acridine orange (AO) 
      and propidium iodide (PI) dyes were used to determine the total cell 
      concentration and microorganism viability, respectively. The results indicated 
      that total cell concentrations determined using FCM were statistically higher 
      (2.62-4.94 times) than those determined using EFM. The difference might be due to 
      cell losses induced by extensive preparations needed for EFM. In addition, EFM 
      and FCM were highly associated for both the total cell concentration and 
      viability. FCM-measured viability was the highest, whereas the culture-measured 
      viability was the lowest. Furthermore, DNA concentrations measured by real-time 
      qPCR with gene probe were highly associated with the total number concentrations 
      measured by either the EFM or FCM. In summary, the three non-culture methods 
      compared here could provide rapid and accurate information about microorganism 
      concentrations and viabilities.
FAU - Chen, Pei-Shih
AU  - Chen PS
AD  - Graduate Institute of Environmental Health, College of Public Health, National 
      Taiwan University, Room 1449, No. 1, Jen Ai Road, 1st Section 100, Taipei, Taiwan 
      R.O.C.
FAU - Li, Chih-Shan
AU  - Li CS
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20050204
PL  - England
TA  - J Environ Monit
JT  - Journal of environmental monitoring : JEM
JID - 100968688
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Bacterial)
RN  - 0 (Fluorescent Dyes)
RN  - 36015-30-2 (Propidium)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Colony Count, Microbial
MH  - DNA Probes
MH  - DNA, Bacterial/analysis
MH  - Escherichia coli/*isolation & purification
MH  - *Flow Cytometry
MH  - Fluorescent Dyes
MH  - *Microscopy, Fluorescence
MH  - *Polymerase Chain Reaction
MH  - Propidium
EDAT- 2005/03/01 09:00
MHDA- 2005/08/02 09:00
CRDT- 2005/03/01 09:00
PHST- 2005/03/01 09:00 [pubmed]
PHST- 2005/08/02 09:00 [medline]
PHST- 2005/03/01 09:00 [entrez]
AID - 10.1039/b415250f [doi]
PST - ppublish
SO  - J Environ Monit. 2005 Mar;7(3):257-62. doi: 10.1039/b415250f. Epub 2005 Feb 4.