No abstract

M ycoplasma genitalium has been implicated as a cause of acute and chronic nonchlamydial nongonococcal urethritis (NCNGU) and pelvic inflammatory disease (PID) (1). In New Zealand, as with many other countries, the preferred treatment for NCNGU is either azithromycin or doxycycline (2). The presence of macrolide resistance in M. genitalium isolates from New Zealand has been reported previously from a small sample size, but there has been no further testing or any continued surveillance reported since then (3). The Microbiology Department in LabPLUS receives specimens mostly from the clinics of the Auckland Regional Sexual Health Service (ARSHS) for M. genitalium testing, which is performed in-house using TaqMan real-time PCR on an ABI 7500 system (Applied Biosystems Life Technologies, NY, USA). This targets the M. genitalium MgPa adhesion gene, as previously described (4). A PlexPCR M. genitalium ResistancePlus kit (SpeeDx Pvt Ltd., Sydney, Australia), a commercial multiplexed real-time PCR assay that detects the presence of M. genitalium and mutations in the 23S rRNA gene (5), was used for this retrospective study. (Ethics approval was received from the Auckland District Health Board [Ethics: 16/CEN/188]). In the study design, M. genitaliumpositive DNA specimens stored at Ϫ70°C from 2009 until 2015 were retrieved and retested by the use of a PlexPCR kit. M. genitalium-positive DNA specimens showing mutations in the 23S rRNA gene by the PlexPCR assay had the 23S rRNA gene sequenced on ABI 3130xl sequencer (Applied Biosystems Life Technologies, NY, USA) to confirm the point mutations in the 23S rRNA gene, using a previously published method (6). Over the period from the start of the M. genitalium testing by LabPLUS in 2009 until 2015, a total of 779 clinical specimens were tested, 149 (19%) of which were positive for M. genitalium. Of the 149 stored M. genitalium-positive samples, 116 (79%) were able to be amplified and further sequenced to confirm the mutation in the 23S rRNA gene detected by PlexPCR. Among the M. genitalium-positive samples recovered for testing by PlexDX PCR, there was agreement in the result with the in-house MgPa real-time assay result. PlexPCR detected mutations in the 23S rRNA gene in 86 of the 116 specimens, all of which were confirmed by in-house sequencing of the 23S rRNA gene. Mutation was detected at both position A2058 (32.5%) and position A2059 (67.5%). The predominant mutants were A2058G and A2059G, while less than 4% of each of A2058C, A2058T, and A2059C were detected. This laboratory-based study shed light on the prevalence of macrolide resistance in the patients presenting with persistent urethritis to the clinics of the ARSHS. Of the 116 specimens tested, 79 were patient specimens sent for testing only once, 17 were patient specimens sent twice within a 1-to 6-month interval to the laboratory

No abstract

UK policy has increasingly promoted early intervention and permanence planning for children who experience, or are at risk of experiencing, abuse or neglect, raising the question of whether these practices have actually increased 'on the ground.' There is already evidence of increased early intervention, in the form of out of home care, in England, as well as Australia and Canada, but thus far we do not know whether early out of home care is increasing in Scotland. Furthermore, there is no research investigating whether rates of permanence planning have changed anywhere in the UK. The current study addressed these gaps through a comparison of two samples of children in Scotland: 110 children born in 2003, and 117 born in 2013, all of whom were placed under compulsory measures of supervision prior to three years of age. The 2013 cohort was significantly more likely than the 2003 cohort to be removed from their parents at birth; to reside away from parents throughout the first three years of life; and to reside away from parents at three years of age. Significantly more of the 2013 cohort than the 2003 cohort had a plan for permanence by three years. These findings are consistent with the view that policy changes in the UK are impacting practice (although practice changes may have resulted from other sources as well / instead). The fall in parental care was largely compensated by an increase in the use of foster care, which has resource implications. Children removed from their parents at birth were usually not returned in the first three years of life, not raised by extended family members, and were separated from one or more siblings. This typically reduced instability for young children, but also entailed substantial birth family fragmentation. The impact on children and families of early removal into foster care must therefore be carefully assessed in light of the increasing prevalence of this practice in Scotland and elsewhere.

M anagement of Mycoplasma genitalium infections is challenging because the limited treatment options have been affected by rapidly evolving resistance to antimicrobial drugs. Molecular approaches are the preferred method of M. genitalium detection, and resistance is determined genotypically. 23S rRNA mutations are associated with macrolide resistance and azithromycin treatment failures (1-3), whereas fluoroquinolone resistance is associated with mutations in the quinolone resistance-determining region, specifically in the gyrA and parC genes (4). Azithromycin is the first-line treatment for M. genitalium infections in New Zealand; second-line treatment relies on moxifloxacin, a fluoroquinolone. A high proportion (72%) of macrolide resistance has been reported in sexual health clinic (SHC) patients in our region (5), and elsewhere in New Zealand fluoroquinolone resistance is reported in 23.3% of M. genitalium-positive specimens from SHC attendees (3), consistent with the high prevalence of macrolide and fluoroquinolone resistance in the Asia-Pacific region (1,3,6). The Study We performed a retrospective study of all specimens referred to the Microbiology Department at Auckland City Hospital (Auckland, New Zealand) for M. genitalium testing in 2017. Referral sites were predominantly general practices in Auckland and SHCs in the Auckland, Northland, and Waikato regions. Ethics approval was granted by the Health and Disability Ethics Committee (approval no. 16/CEN/188). DNA had been extracted from specimens following a diagnostic workflow and stored at −80°C. We retrieved DNA samples that tested positive for M. genitalium using real-time PCR (5,7) for this study. We detected 23S rRNA mutations at nucleotide positions 2058 and 2059 (Escherichia coli numbering) by using the commercially available PlexPCR kit Resistance Plus MG (SpeeDx, https://plexpcr.com) (5) and gyrA and parC mutations by using PCR amplification of M. genitalium nucleotides 172-402 of gyrA and 164-483 of parC (8), followed by sequencing on the Applied Biosystems 3130xl sequencer (Life Technologies, https://www.thermofisher.com). Sequences were aligned against M. genitalium reference genes (Gen-Bank accession nos. CP003773 for gyrA and parC for U25549) by using SeqMan II (DNASTAR, https:// www.dnastar.com) and the mutations reported by using M. genitalium numbering. We compared prevalence of resistance-associated mutations in community and SHC cohorts by using a χ 2 test (α = 5%). We tested 302 clinical specimens from 247 patients; 33% (101/302) of samples from 34% (84/247) of patients were M. genitalium DNA-positive. Four samples from 3 patients were excluded from subsequent analyses because insufficient PCR products were obtained for sequencing. We used the remaining 97