Novel real-time PCR assay for simultaneous detection and differentiation of Clostridium chauvoei and Clostridium septicum in clostridial myonecrosis.
A real-time PCR assay based on the 16S rRNA gene sequence was designed for differentiation of blackleg-causing Clostridium chauvoei and Clostridium septicum, a phylogenetically closely related bacterium responsible for malignant edema. In order to exclude false-negative results, an internal amplification control was included in the assay. A set of three probes, one specific for C. chauvoei, one specific for C. septicum, and one specific for both species, permitted unequivocal detection of C. chauvoei in tests of 32 Clostridium sp. strains and 10 non-Clostridium strains. The assay proved to be sensitive, detecting one genome of C. chauvoei or C. septicum per PCR and 1.79 x 10(3) C. chauvoei cells/g artificially contaminated muscle tissue. In tests of 11 clinical specimens, the real-time PCR assay yielded the same results as an established conventional PCR method.
Department of Large Animals and Veterinary Public Health, University of Veterinary Medicine Vienna, Veterinaerplatz 1, A-1210 Vienna, Austria. ingeborg.hein@vu-wien.ac.at
Anna
A
Halm
Martin
M
Wagner
Josef
J
Köfer
Ingeborg
I
Hein
D003015:Clostridium Infections / Q000175:diagnosis* / Q000382:microbiology; D046975:Clostridium chauvoei / Q000145:classification* / Q000302:isolation & purification*; D052557:Clostridium septicum / Q000145:classification* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012015:Reference Standards; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S