CLINICAL REPORT
1235 Advances in dermatology and venereology ActaDV Acta Dermato-Venereologica ActaDV
Mycoplasma genitalium and Macrolide Resistance-associated Mutations in the Skåne Region of Southern Sweden 2015
Ola FORSLUND 1, Max HJELM 1, Raghd EL-ALI 1, Annika JOHNSSON 2 and Carina BJARTLING ²
1
Department of Laboratory Medicine, Division of Medical Microbiology, Lund University, Skåne Laboratory Medicine, and 2 Centre for Sexual Health, Skåne University Hospital, Lund, Sweden
Mycoplasma genitalium is a sexually transmitted infection ordinarily treated with azithromycin. Emerging resistance to macrolide is linked to mutations in the 23S rRNA gene. We analysed the frequency of such mutations of M. genitalium isolates from patients that were symptomatic, and from sexual partners of symptomatic individuals, from October to December of 2015, in the Skåne Region of Sweden. Mutations were analysed by the use of DNA sequencing. Overall, 11.9 %( 145 / 1,311) and 17.0 %( 116 / 704) of females and males were positive for M. genitalium, respectively. Macrolide resistant mutations were detected in 13 %( 31 / 239) of M. genitalium isolates from first-test patient samples. Twenty-one( 8.8 %) and 10( 4.2 %) of the isolates had point mutations of the 23S-gene at position 2072 and 2071, respectively. Two different M. genitalium isolates were detected simultaneously in two cases. In summary, we found a relatively low rate of macrolide-resistant M. genitalium in the region of Southern Sweden.
Key words: mycoplasma genitalium; macrolide; azithromycin; resistance.
Accepted Jul 5, 2017; Epub ahead of print Jul 6, 2017 Acta Derm Venereol 2017; 97: 1235 – 1238.
Corr: Ola Forslund, Department of Laboratory Medicine, Division of Medical Microbiology, Lund University, Skåne Laboratory Medicine, Sölvegatan 23, SE-222 53 Lund, Sweden. E-mail: ola. forslund @ med. lu. se
Mycoplasma genitalium was discovered from men with non-gonococcal urethritis( NGU) in 1981( 1). M. genitalium is a sexually transmitted bacterium and is a major cause of NGU in males( 2). It is also associated with female cervicitis and complications such as pelvic inflammatory disease( PID)( 3). The macrolide azithromycin is commonly used for treatment of M. genitalium. Mutations in region V of the 23S rRNA gene of M. genitalium are consistently linked to macrolide resistance and were first described in 2008 by Jensen et al.( 4). However, emerging resistance problem against M. genitalium have been reported in Scandinavia, where a Swedish study observed an increase in macrolide resistance before introduction of treatment from 0 % in 2006 to 18 % in 2011( 5), and a Denmark survey detected macrolide resistance in 38 % of M. genitalium-positive unselected patients( 6). Recently, a Swedish study reported macrolide resistance of 18 % among M. genitalium isolates collected throughout 2012 in the Stockholm region( 7). The aim of the present study was to investigate the prevalence of macrolide-associated mutations among M. genitalium isolates in the Skåne Region within the 4 th quarter of 2015.
MATERIALS AND METHODS Study population
Within the region of Skåne Sweden with 1.3 million habitants 1 urogenital samples, from 3,167 males( mean age 32 years, median 29 years, range 13 – 82 years) and 5,636 women( mean age 28 years, median 26 years, range 2 – 77 years) who were seeking care at diverse clinics, such as sexual health clinics, youth clinics and general practitioners, were routinely tested for M. genitalium during 2015. According to Swedish guidelines, urogenital samples sent for M. genitalium diagnostics were from patients with clinical symptoms related to M. genitalium and from asymptomatic sexual partners to M. genitalium positive patients. The samples were analysed for M. genitalium at the Clinical Microbiology Laboratory, Region Skåne, Sweden by the use of a real-time PCR( 8).
Study design
A retrospective study of macrolide-resistance was performed on M. genitalium isolates collected from October to December 2015. The M. genitalium isolates were originally detected in urogenital samples by routine testing by the use of real-time PCR( 8). A sample was classified as a first-test patient sample if no previous sample had been collected within the preceding 6 weeks. The length of 6 weeks was chosen in order to include relatively recently acquired infections among the first-test patient samples. This period is in agreement with that of the new European guidelines from 2016 where samples for test of cure should be collected no earlier than 3 weeks after start of treatment( 9).
Laboratory procedures
From samples positive for M. genitalium, remaining materials of nucleic acid extractions( Magna LC, Total NA. Large Volume kit, 500 ul input 50 ul output) from APTIMA Urine / Vaginal / Swab collection tubes were stored at – 20 ° C. For amplification of the M. genitalium 23S rRNA gene of region V, we used a 20 µ l reaction of 1 x Perfecta™ qPCR Fast Mix™ Low Rox,( Quanta Biosciences, Gaithersburg, Maryland, USA), 0.125 %( w / v) BSA( Ultrapure™, Ambion, Thermofisher Scientific, MA, USA), 0.2 µ m of each forward( Mg 23S-1992F) and reverse primer( Mg 23S-2138R)( 4)( LGC Biosearch Technologies, Risskov, Denmark) and 5 µ l of nucleic acid extraction from each sample. The PCR reaction was carried out in an automated thermocycler( ABI 9700) as follows; 20’’ at 95 ° C and then 50 cycles of 3’’ at 95 ° C and 1’ at 60 ° C. Purified DNA of M. genitalium( Amplirun ® Mycoplasma genitalium
1
“ Population in the country, counties and municipalities, 31 December 2015”. Central Bureau of Statistics. Read February 23, 2016.
This is an open access article under the CC BY-NC license. www. medicaljournals. se / acta Journal Compilation © 2017 Acta Dermato-Venereologica. doi: 10.2340 / 00015555-2746 Acta Derm Venereol 2017; 97: 1235 – 1238