| Summary: | <i>Mycoplasma (M.) parvum</i> is a hemotrophic bacterium circulating in the blood of pigs but is not considered a primary pathogen. Only a handful of studies dealing with this agent have been published since its first description in 1951, and many issues, including epidemiology and the impact of subclinical infections, are yet to be elucidated. This study aimed to establish a <i>M. parvum</i> specific real-time PCR for its detection and quantification in porcine blood and the application of this assay to obtain insights into the occurrence of <i>M. parvum</i> in German pigs. Furthermore, 16S rDNA amplicons of <i>M. parvum</i> positive blood samples were phylogenetically analyzed using MEGA 11 software. The established qPCR targeting the <i>M. parvum</i> glyceraldehyde-3-phosphate dehydrogenase encoding gene (<i>gap</i>) showed a lower detection limit of 10 gene copies per reaction and no cross-reactivity within the specificity test. A total of 36.0% (<i>n</i> = 72) of the sampled fattening pigs, 25.0% (<i>n</i> = 15) of the sows, and 4.37% (<i>n</i> = 8) of the boars tested <i>M. parvum</i> positive. The dendrogram showed the typical allocation of the <i>M. parvum</i> isolates into the “haemominutum group” subgroup within the hemotrophic <i>Mycoplasma</i> species. Both the novel established qPCR and the obtained epidemiological data can serve as an important basis for future studies dealing with <i>M. parvum.</i>
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