Lindroth Ylva, Hansson Lucia, Forslund Ola
Clinical Microbiology, Infection Prevention and Control, Office for Medical Services, Lund, Sweden.
Department of Translational Medicine, Lund University, Malmö, Sweden.
APMIS. 2025 Jan;133(1):e13477. doi: 10.1111/apm.13477. Epub 2024 Oct 11.
Azithromycin, a macrolide antibioticum, is the first-line treatment for Mycoplasma genitalium (MG), but resistant MG is an increasing problem. Macrolide resistance-mediated mutations (MRM) has been linked to point mutations in region V of the MG 23S rRNA gene. We have evaluated an open access analyzer (Panther Fusion, Hologic Inc) for detectability of MRM (mutations A2071G and A2072G) and MG wild type (WT) in clinical samples. Also, the agreement of the Panther Fusion assay results with a corresponding established In-house MRM-WT PCR (ABI 7500) was calculated. Left over material from 55 clinical samples positive for MG by the Aptima test (Hologic) based on transcription-mediated amplification (TMA), collected from January to February 2023 in Region Skåne, Sweden, was analyzed. Specific amplification curves were generated for positive controls of MG mutations (A2071G and A2072G) and WT by the Panther Fusion assay. The limit of detection (LOD) was 5.3 copies/mL for WT, 8.1 copies/mL for mutation A2071G, and 81 copies/mL for mutation A2072G. The overall concordance was 91% between the Panther Fusion and the In-house PCR (Kappa 0.621, 95% CI; 0.327-0.914) for detection of WT or MRM in MG-positive clinical samples. The Panther Fusion detected MRM in 20% (11/55) and WT in 62% (34/55) of the samples. The corresponding In-house PCR results were 25% (14/55) and 65% (36/55). In summary, the Panther Fusion assay demonstrated detection of low copy number of MRM and WT of MG. Among clinical samples substantial agreement between the Panther Fusion and In-house PCR results was observed. Integrating MG-analysis (TMA) and MRM-WT assay on the Panther platform could make MRM testing more readily available. However, the Panther Fusion had a lower success rate (82% vs 90%) for macrolide susceptibility testing, hence testing with a complementary method should be considered for samples where neither WT nor MRM MG are detectable.
阿奇霉素是一种大环内酯类抗生素，是生殖支原体（MG）的一线治疗药物，但MG耐药问题日益严重。大环内酯类耐药介导的突变（MRM）与MG 23S rRNA基因V区的点突变有关。我们评估了一种开放式分析仪（Panther Fusion，Hologic公司）对临床样本中MRM（A2071G和A2072G突变）和MG野生型（WT）的检测能力。此外，还计算了Panther Fusion检测结果与相应的已建立的内部MRM-WT PCR（ABI 7500）结果的一致性。分析了2023年1月至2月在瑞典斯科讷地区通过基于转录介导扩增（TMA）的Aptima检测（Hologic）检测为MG阳性的55份临床样本的剩余材料。通过Panther Fusion检测对MG突变（A2071G和A2072G）和WT的阳性对照产生了特异性扩增曲线。WT的检测限（LOD）为5.3拷贝/毫升，A2071G突变为8.1拷贝/毫升，A2072G突变为81拷贝/毫升。在MG阳性临床样本中检测WT或MRM时，Panther Fusion与内部PCR的总体一致性为91%（Kappa 0.621，95%CI；0.327-0.914）。Panther Fusion在20%（11/55）的样本中检测到MRM，在62%（34/55）的样本中检测到WT。相应的内部PCR结果分别为25%（14/55）和65%（36/55）。总之，Panther Fusion检测证明可检测到低拷贝数的MG的MRM和WT。在临床样本中，观察到Panther Fusion与内部PCR结果之间有高度一致性。在Panther平台上整合MG分析（TMA）和MRM-WT检测可以使MRM检测更容易获得。然而，Panther Fusion进行大环内酯类药物敏感性检测的成功率较低（82%对90%），因此对于既未检测到WT也未检测到MRM MG的样本，应考虑用互补方法进行检测。
