Rapid detection and identification of Clostridium chauvoei by PCR based on flagellin gene sequence.
We developed a one-step polymerase chain reaction (PCR) system that specifically detects Clostridium chauvoei. Oligonucleotide primers were designed to amplify a 516-bp fragment of the structural flagellin gene. The specificity of the PCR was investigated by analyzing 59 strains of clostridia, and seven strain of other genera. A 516-bp fragment could be amplified from all the C. chauvoei strains tested, and no amplification was observed by using DNAs from the other strains tested, including Clostridium septicum. Similarly, this PCR-based method specifically detected C. chauvoei DNA sequences in samples of muscle and exudate of obtained from mice within 12h of inoculation. In tests using samples of muscle or liver, the limit of detection was about 200 organisms per reaction. These results suggest that the one-step PCR system may be useful for direct detection and identification of C. chauvoei in clinical specimens.
Kojima
A
A
National Veterinary Assay Laboratory, Ministry of Agriculture, Forestry and Fisheries, 1-15-1, Tokura, Kokubunji, 185-8511, Tokyo, Japan. kojima@nval.go.jp
Uchida
I
I
Sekizaki
T
T
Sasaki
Y
Y
Ogikubo
Y
Y
Tamura
Y
Y
D000818:Animals; D001681:Biological Assay; D003013:Clostridium / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D003015:Clostridium Infections / Q000175:diagnosis* / Q000382:microbiology; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D005260:Female; D005408:Flagellin / Q000737:chemistry / Q000235:genetics*; D008099:Liver / Q000382:microbiology; D051379:Mice; D018482:Muscle, Skeletal / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D005408:Flagellin