To determine the presence and genotypic macrolide susceptibility of Mycoplasma amphoriforme, and presence of Ureaplasma spp., and Mycoplasma fermentans among clinical samples from England previously investigated for Mycoplasma pneumoniae.
Quantitative and conventional PCR were used to retrospectively screen a collection of 160 clinical samples, previously submitted to Public Health England (PHE) for detection of M. pneumoniae, between October 2016 and December 2017. Samples which were positive for M. amphoriforme DNA were further investigated for mutations associated with genotypic macrolide resistance by sequencing of domain V of the 23s rRNA.
Mycoplasma amphoriforme were detected in 10/160 (6.3%) samples, Ureaplasma parvum were detected in 4/160 samples (2.5%) with 0/160 M. fermentans detections. Of the nine individuals (two samples were from the same patient) in which M. amphoriforme were detected, eight were male (10 – 60 years age range) and one was female (30 – 40 years age range). One individual, with cystic fibrosis was positive for both M. amphoriforme and U. parvum. All M. amphoriforme DNA were genotypically susceptible for macrolides.
Mycoplasma amphoriforme were found in clinical samples including the lower respiratory tract samples of patients with pneumonia. In the absence of other respiratory pathogens, these data suggest a potential role of this organism in human disease, with no evidence of acquired macrolide resistance. Ureaplasma parvum was detected in cerebrospinal fluid and respiratory tract samples. These data suggest the need to consider these atypical respiratory pathogens in future diagnostic investigations.

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