Phylogenetic analysis and PCR detection of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum based on the flagellin gene.
The flagellin genes (fliC) of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene (fliC) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I (), are more closely related to Bacillus subtilis than to Clostridium difficile, which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei, C. haemolyticum, C. novyi types A and B, and C. septicum. PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia.
Sasaki
Yoshimasa
Y
National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1 Tokura, Kokubunji, Tokyo, Japan. sasakiy@nval.go.jp
Kojima
Akemi
A
Aoki
Hiroshi
H
Ogikubo
Yasuaki
Y
Takikawa
Noriyasu
N
Tamura
Yutaka
Y
D001483:Base Sequence; D003001:Cloning, Molecular; D003013:Clostridium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D005408:Flagellin / Q000235:genetics*; D005784:Gene Amplification; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D013045:Species Specificity
D004275:DNA, Ribosomal; D005408:Flagellin