Universal primer PCR with DGGE for rapid detection of bacterial pathogens.
A universal primer PCR (UPPCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens. The results show that this method is efficient at amplifying the conserved regions of bacterial 16S rRNA genes with universal primers and can detect causative bacterial pathogens rapidly. Six species of bacteria from fisheries (Pseudomonas fluorescens, Vibrio anguillarum, Aeromonas hydrophila, Vibrio fluvialis, Providencia rettgeri and Aeromonas sobria) were examined. Our results indicate that the approach we undertook can be adopted not only for axenic bacterial populations but also for mixed communities as well. Furthermore, we were able to achieve the rapid detection of multiple bacteria a single in sample. In addition, UPPCR-DGGE was shown to be better than previously reported UPPCR-single-stranded conformation polymorphism (SSCP)-based methods for the rapid detection of bacterial pathogens.
Ji
Niannian
N
Department of Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, PR China.
Peng
Bo
B
Wang
Guizhong
G
Wang
Sanying
S
Peng
Xuanxian
X
D000818:Animals; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary*; D005393:Fish Diseases / Q000382:microbiology*; D005399:Fishes; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000254:growth & development*; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S