Phylogenetic analysis and PCR detection of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum based on the flagellin gene

Successful Treatment of Bacillary Hemoglobinuria in Japanese Black Cows

“…The presence of the fliC gene was also confirmed by nested PCR using the forward primer 5- AAGAGGGCTTAATCAAGCTTCAA-3 (nucleotide positions 191-213) and the reverse primer 5-TGAATAT-TATTTCCACTTTCTCC-3 (nucleotide positions 666-688), as well as nucleotide sequencing according to the general protocol (Hokkaido System Science, Sapporo, Japan). The nucleotide sequence (452 base pairs) of the second PCR primers corresponded to the one reported previously (nucleotide positions 214-665) [8]. This result demonstrated that patient 3 was infected with C. haemolyticum and, hence, the diagnosis of BHU was established.…”

“…haemolyticum detection in the blood samples of patient 3 and 4 was conducted by using the PCR system based on the flagellin gene (fliC) sequence (DDBJ accession no. AB058939) reported by Sasaki et al [8]. The sample from patient 3 was collected before initiation of the treatment, i.e.…”

“…It has been reported that isolation of C. haemolyticum is time-consuming and very difficult, since it requires strict anaerobic conditions, and that clinical specimens are often contaminated with other anaerobic bacteria, including environmental clostridia from soil, which grow faster than the pathogenic bacteria in the culture medium [9,11]. Recently, Sasaki et al [8] reported the possibility of rapid identification of pathogenic clostridia with the help of the established multiplex PCR system. We definitively diagnosed patient 3 with BHU on the basis of the PCR and sequencing results.…”

Fatal <i>Clostridium novyi</i> Type B Infection in a Sow

“…To discriminate between Clostridium species, a PCR amplifying the flagellin gene fliC, present on the chromosome, was also performed as previously described (Sasaki et al 2002). This PCR, using one forward and five reverse primers, has been developed to identify and differentiate between C. novyi types A and B, C. haemolyticum, Clostridium chauvoei and Clostridium septicum (Sasaki et al 2002).…”

“…They can be divided into four types based on toxin production (Sasaki et al 2002). The α toxin is produced by types A and B, and the β toxin by types B and D (C. novyi type D is also called Clostridium haemolyticum) (Sasaki et al 2002).…”