Acholeplasma laidlawii PG8 ultramicroforms amplificate selectively rrnB nucleotide sequences

Abstract

Mycoplasmas are frequent contaminants of in vitro animal cell cultures. Despite a broad spectrum of modern methods, detection of mycoplasmas remains a serious problem. The situation is complicated by the fact that mycoplasmas may be presented in cell cultures or biological samples by viable but unculturable forms (ultramicroforms). We found that the DNA of Acholeplasma laidlawii PG8 ultramicroforms showed selective amplification of the rrnB nucleotide sequences while vegetative cells of the mycoplasma showed amplification both for rrnA and rrnB sequences. The role of enzyme deproteinization in PCR results was also shown. The results presented in this report indicate that the optimisation of primer sequences as well as PCR regime may be crucial steps in detection and differentiation of vegetative forms and ultramicroforms of A. laidlawii.