PMID- 21791228
OWN - NLM
STAT- MEDLINE
DCOM- 20111216
LR  - 20131121
IS  - 1872-8359 (Electronic)
IS  - 0167-7012 (Linking)
VI  - 87
IP  - 1
DP  - 2011 Oct
TI  - Development of multiplex PCR assay for rapid detection of Riemerella 
      anatipestifer, Escherichia coli, and Salmonella enterica simultaneously from 
      ducks.
PG  - 64-9
LID - 10.1016/j.mimet.2011.07.007 [doi]
AB  - Three pathogens, Riemerella anatipestifer, Escherichia coli, and Salmonella 
      enterica, are leading causes of bacterial fibrinous pericarditis and 
      perihepatitis in ducks in China and worldwide. It is difficult to differentiate 
      these pathogens when obtaining a diagnosis on clinical signs and pathological 
      changes. The aim of this research was to develop a multiplex polymerase chain 
      reaction (m-PCR) that could discriminate R. anatipestifer, E. coli, and S. 
      enterica rapidly in field isolates, or detect the three bacteria in clinical 
      samples from diseased ducks. We selected the DnaB helicase (dnaB) gene of R. 
      anatipestifer, alkaline phosphatase (phoA) gene of E. coli and invasion protein 
      (invA) gene of S. enterica as target genes. In optimized conditions, the 
      limitation of detection was approximately 10(3) colony forming units (CFU) of 
      each of these three bacterial pathogens per PCR reaction tube. The m-PCR method 
      showed specific amplification of respective genes from R. anatipestifer, E. coli, 
      and S. enterica. Using the m-PCR system, bacterial strains isolated from diseased 
      ducks in our laboratory were categorized successfully, and the pathogens could 
      also be detected in clinical samples from diseased ducks. Therefore, the m-PCR 
      system could distinguish the three pathogens simultaneously, for identification, 
      routine molecular diagnosis and epidemiology, in a single reaction.
CI  - Copyright (c) 2011 Elsevier B.V. All rights reserved.
FAU - Hu, Qinghai
AU  - Hu Q
AD  - Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural 
      Sciences (CAAS), 518 Ziyue Road, Shanghai 200241, PR China.
FAU - Tu, Jing
AU  - Tu J
FAU - Han, Xiangan
AU  - Han X
FAU - Zhu, Yinyu
AU  - Zhu Y
FAU - Ding, Chan
AU  - Ding C
FAU - Yu, Shengqing
AU  - Yu S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110721
PL  - Netherlands
TA  - J Microbiol Methods
JT  - Journal of microbiological methods
JID - 8306883
RN  - 0 (Bacterial Proteins)
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (Genetic Markers)
RN  - 147652-43-5 (invA protein, Bacteria)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
RN  - EC 3.1.3.1 (phoA protein, E coli)
RN  - EC 3.6.4.12 (DnaB Helicases)
SB  - IM
MH  - Alkaline Phosphatase/genetics
MH  - Animals
MH  - Bacterial Proteins/genetics
MH  - Bird Diseases/*microbiology
MH  - Colony Count, Microbial
MH  - DnaB Helicases/genetics
MH  - Ducks/*microbiology
MH  - Escherichia coli/genetics/growth & development
MH  - Escherichia coli Infections/microbiology/*veterinary
MH  - Escherichia coli Proteins/genetics
MH  - Flavobacteriaceae Infections/microbiology/*veterinary
MH  - Genetic Markers
MH  - Hepatitis, Animal/microbiology
MH  - Limit of Detection
MH  - Molecular Typing/*methods
MH  - Multiplex Polymerase Chain Reaction/*methods
MH  - Pericarditis/microbiology/veterinary
MH  - Peritonitis/microbiology/veterinary
MH  - Riemerella/genetics/growth & development
MH  - Salmonella Infections, Animal/*microbiology
MH  - Salmonella enterica/genetics/growth & development
EDAT- 2011/07/28 06:00
MHDA- 2011/12/17 06:00
CRDT- 2011/07/28 06:00
PHST- 2011/04/07 00:00 [received]
PHST- 2011/06/28 00:00 [revised]
PHST- 2011/07/08 00:00 [accepted]
PHST- 2011/07/28 06:00 [entrez]
PHST- 2011/07/28 06:00 [pubmed]
PHST- 2011/12/17 06:00 [medline]
AID - S0167-7012(11)00257-0 [pii]
AID - 10.1016/j.mimet.2011.07.007 [doi]
PST - ppublish
SO  - J Microbiol Methods. 2011 Oct;87(1):64-9. doi: 10.1016/j.mimet.2011.07.007. Epub 
      2011 Jul 21.