A novel polymerase chain reaction assay for the detection of seven Mycoplasma species of cattle origin

Abstract

The aim of this study was to develop a pair of primers for detecting ruminant mycoplasma pathogens.We designed a set of primers based on the most similar sequences within 16sRNA regions of seven Mycoplasma spp. These primers have high sensitivity for detecting Mycoplasma dispar, M. arginine, M. canadense, M. bovis, M. alkalescens, M. californicum, and M. bovisgenitalium within the annealing temperature range of 46°C to 48°C. The minimum amount of DNA that can be detected using the protocol is 250 ng, which is equivalent to 2,000 cfu/mL. The primers can detect mycoplasma from DNA extracted directly from milk samples. The common bovine mastitis pathogens of Staphylococcus aureus coagulase-negative staphylococci, Escherichia coli, Streptococcus uberis, Klebiella pneumonia, Korucia rosea, and Acholeplasma spp. were not detected by the primers. We believe the high sensitivity and speci city of these primers make them useful for detecting infection with seven Mycoplasma species in ruminants, allowing the primers to be used in clinical settings.