PMID- 19566721
OWN - NLM
STAT- MEDLINE
DCOM- 20100420
LR  - 20091216
IS  - 1365-2672 (Electronic)
IS  - 1364-5072 (Linking)
VI  - 108
IP  - 1
DP  - 2010 Jan
TI  - Comparative study of a new quantitative real-time PCR targeting the 
      xylulose-5-phosphate/fructose-6-phosphate phosphoketolase bifidobacterial gene 
      (xfp) in faecal samples with two fluorescence in situ hybridization methods.
PG  - 181-93
LID - 10.1111/j.1365-2672.2009.04408.x [doi]
AB  - AIMS: To detect and enumerate bifidobacteria in faeces with a new quantitative 
      multiplex real-time PCR (qPCR) method and to compare the results obtained with 
      fluorescence in situ hybridization (FISH) methods. METHODS AND RESULTS: A 
      multiplex qPCR assay was developed, which enabled the enumeration of 
      Bifidobacterium spp. by targeting the bifidobacterial 
      xylulose-5-phosphate/fructose-6-phosphate phosphoketolase gene (xfp) and total 
      bacteria using universal Eub-primers targeting 16S rRNA gene from the domain 
      bacteria. The qPCR assay showed high sensitivity and specificity and a low 
      detection limit of about 2.5 x 10(3) bifidobacterial cells per gram of faeces. 
      The qPCR results were compared with FISH combined with microscopy or flow 
      cytometry (FCM). No statistical differences among bifidobacterial counts averages 
      measured in adult faeces with the three methods were observed. Total bacterial 
      count averages were higher with the FISH method coupled with microscopic analyses 
      compared to FISH with FCM, whereas total cell numbers estimated by qPCR were 
      intermediate between the two FISH methods. CONCLUSIONS: The new qPCR assay was 
      shown to be sensitive, rapid and accurate for enumerating bifidobacteria in 
      faeces. SIGNIFICANCE AND IMPACT OF THE STUDY: This method is a valuable 
      alternative for other molecular methods for detecting faecal bifidobacteria, 
      especially when their counts are below the detection limit of the FISH methods.
FAU - Cleusix, V
AU  - Cleusix V
AD  - Institute of Food Science and Nutrition, Swiss Federal Institute of Technology, 
      Zentrum, Zurich, Switzerland.
FAU - Lacroix, C
AU  - Lacroix C
FAU - Dasen, G
AU  - Dasen G
FAU - Leo, M
AU  - Leo M
FAU - Le Blay, G
AU  - Le Blay G
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Appl Microbiol
JT  - Journal of applied microbiology
JID - 9706280
RN  - 0 (Bacterial Proteins)
RN  - EC 4.1.2.- (Aldehyde-Lyases)
RN  - EC 4.1.2.22 (fructose-6-phosphate phosphoketolase)
SB  - IM
MH  - Adult
MH  - Aldehyde-Lyases/*genetics
MH  - Bacterial Proteins/*genetics
MH  - Bifidobacterium/enzymology/*genetics
MH  - Feces/*microbiology
MH  - Female
MH  - Flow Cytometry/methods
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Infant
MH  - Male
MH  - Middle Aged
MH  - Polymerase Chain Reaction/*methods
MH  - Sensitivity and Specificity
EDAT- 2009/07/02 09:00
MHDA- 2010/04/21 06:00
CRDT- 2009/07/02 09:00
PHST- 2009/07/02 09:00 [entrez]
PHST- 2009/07/02 09:00 [pubmed]
PHST- 2010/04/21 06:00 [medline]
AID - JAM4408 [pii]
AID - 10.1111/j.1365-2672.2009.04408.x [doi]
PST - ppublish
SO  - J Appl Microbiol. 2010 Jan;108(1):181-93. doi: 10.1111/j.1365-2672.2009.04408.x.