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Successful sequencing (99%) of the cdtA and cdtb genes was used to confirm the identity of one isolate that was positive for the binary toxin genes (Macrogen, Korea).
The presence of the cadF, cdtA, cdtB, and cdtC genes plus that of the full cdt cluster and the iam marker was determined by PCR  (Table 1).
Number Serotype * Virulence marker Resistance marker 1 O103:H7 hemL, lpfA, tsh, iroN, iss 2 On.d.:H19 blaTEM 3 O103:H7 hemL, lpfA, tsh, iroN, iss 4 O103:H7 IpfA, tsh, iroN, iss 5 On.d.:H38 stx2 (subtype 2a or 2c or 2d), ehxA, espI, espP, saa, iha, subA, iss 6 On.d.:H2 cba, cdtB, cma, cnf1, f17-G, IpfA, iroN, iss 7 On.d.:H31 IpfA, cdtB, cnf1, f17-A, f17-G 8 O103:H7 hemL, lpfA, tsh, tetA iroN, iss 9 On.d.:H49 hemL, lpfA 10 On.d.:H2 IpfA, cba, cdtB, cma, cnf1, f17-G, iro, iss 11 O103:H7 hemL, lpfA, tsh, tetA iroN, iss 12 On.d.:Hn.d.
Cdt is an AB type toxin, with the specific receptor subunits named CdtA and CdtC, and the cytotoxic component designated CdtB. CdtB exhibits DNase I activity, disrupting the phosphodiester bonds in chromosomal DNA.
dfor=(cdtb/2*((atb*rhow)*vel2)) where (cdtb) is the drag coefficient, (rhow) is the density of seawater, and (atb) is a characteristic area of the towed body.
After anaerobic incubation at 37[degrees]C for 72 hours, all colonies were subjected to thermal DNA extraction (BAUMS et al., 2014), and then, multiplex PCR was used to detect a housekeeping gene (tpi), toxins A (tcdA) and B (tcdB) and a binary toxin gene (cdtB) as previously described (SILVA et al., 2011).
[10.] Samie A, Obi CL, Franasiak J, et al PCR detection of Clostridium difficile triose phosphate isomerase (tpi), Toxin A (tcdA), Toxin B (tcdB), Binary Toxin (cdtA, cdtB), and tcdC genes in Vhembe District, South Africa.
We further examined the intimin subtype by sequencing the eae gene, the chromosome integration site of the locus of enterocyte effacement encoding the eae gene and a set of type III secretion system genes, and the presence and subtype of the cdtB gene as described (5).
CDT is encoded by three adjacent genes, cdtA, cdtB and cdtC.
In addition to confirming identity, most techniques also detect the genes encoding toxin A (tcdA), toxin B (tcdB) and additional virulence factors such as the binary toxin (cdtB) (SILVA et al., 2011).
The presence of binary toxin gene (cdtB) and deletion in the negative regulator of the pathogenecity locus, tcdC, were determined by using Cepheid GeneXpert PCR (Cepheid, Sunnyvale, CA, USA).
difficile was isolated from both foals, and the two strains were positive by PCR for the toxin A (tdcA) and B (tcdB) genes but negative for the binary toxin gene (cdtB).
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