A PCR scheme for differentiation of organisms belonging to the Mycoplasma mycoides cluster

Comparison of the 16S-23S rRNA intergenic spacer regions among strains of the Mycoplasma mycoides cluster, and reassessment of the taxonomic position of Mycoplasma sp. bovine group 7.

“…The CAP-21 genomic region was used by Taylor et al (1992) to design a DNA probe identifying M. mycoides subspecies and also a PCR assay for identification of the SC type (Bashiruddin et al, 1994). Hotzel et al (1996) developed a PCR system for differentiation of all cluster groups which exploited nucleotide sequence variations in the same region. Other PCR approaches included RFLP analysis of the amplified CAP-21 region (Rodriguez et al, 1997) and arbitrarily primed amplification (Rawadi et al, 1995).…”

“…Mycoplasma DNA was extracted from 100 ml culture (10( c.f.u. ml −" ) as described previously (Hotzel et al, 1996). Amplification of the 16S-23S rRNA intergenic spacer region was performed according to the method described elsewhere (Harasawa, 1999) by using primers F (5h-CCCGTC-ACACCATGAGAGTT-3h) and R (5h-TCGGCTCC-ATTTTCCAAGGC-3h) for amplification, with denaturation at 94 mC for 30 s, annealing at 55 mC for 100 s and extension at 72 mC for 100 s in a TSR-300 thermal cycler (Iwaki Glass).…”

Monoclonal Antibody Differentiation of Mycoplasma mycoides subsp. mycoides Small-Colony Strains Causing Contagious Bovine Pleuropneumonia from Less Important Large-Colony Strains

“…The three MmmLC strains and the two M. mycoides subsp. capri strains included in Table 1 have been notoriously difficult to differentiate from MmmSC strains by other assays (10,18). All of the Mycoplasma strains that were not inhibited by MAb PK-2 in Table 1 were inhibited by either polyclonal antiserum to each of the five Mycoplasma groups (SC, LC, capri, capricolum, and BSG7) or a MAb to the M. capricolum subsp.…”

Genotypic Methods for Diagnosis of Mycoplasmal Infections in Humans, Animals, Plants and Cell Cultures

“…mycoidessubsp. inycoidesSC (Bashiruddin et al, 1994;Dedieu etal., 1994a;Hotzel et al, 1996). Bashiruddin et al (1994) used the CAP-21 probe sequence to design PCR primers that amplify all the members of theM.…”