改进淋病奈瑟菌拓扑异构酶 A 基因型的测定方法可用于临床标本。
Improved determination of Neisseria gonorrhoeae gyrase A genotype results in clinical specimens.
机构信息
Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA.
Department of Medicine, Boston Children's Hospital, Boston, MA, USA.
出版信息
J Antimicrob Chemother. 2019 Oct 1;74(10):2913-2915. doi: 10.1093/jac/dkz292.
BACKGROUND
The emergence of drug-resistant Neisseria gonorrhoeae has prompted the development of rapid molecular assays designed to determine antimicrobial susceptibility. One common assay uses high-resolution melt analysis to target codon 91 of the gyrase A gene (gyrA) to predict N. gonorrhoeae susceptibility to ciprofloxacin.
METHODS
We extracted DNA from remnant clinical specimens that had previously tested positive for N. gonorrhoeae using the Aptima Combo 2 for CT/NG assay (Hologic, San Diego, CA, USA). We selected DNA extracts from specimens with indeterminate, WT and mutant gyrA genotype results from a previous study using high-resolution melt analysis to detect the gyrA codon 91 mutation. We re-tested those specimens using the recently CE-marked ResistancePlus GC (beta) assay (SpeeDx, Sydney, Australia).
RESULTS
Of 86 specimens with indeterminate gyrA genotypes on high-resolution melt analysis, the ResistancePlus GC (beta) assay (SpeeDx) identified 30 (35%) WT, 22 (26%) mutant and 34 (40%) indeterminate gyrA genotypes.
CONCLUSIONS
The ResistancePlus GC (beta) assay showed improved N. gonorrhoeae gyrA genotype determination compared with a prior gyrA genotypic high-resolution melt assay.
背景
耐抗生素淋病奈瑟菌的出现促使人们开发了旨在确定抗菌药物敏感性的快速分子检测方法。一种常见的检测方法使用高分辨率熔解分析来靶向拓扑异构酶 A 基因（gyrA）的密码子 91，以预测淋病奈瑟菌对环丙沙星的敏感性。
方法
我们从先前使用 Aptima Combo 2 用于 CT/NG 检测（美国圣地亚哥 Hologic）检测为淋病奈瑟菌阳性的剩余临床标本中提取 DNA。我们从先前使用高分辨率熔解分析检测 gyrA 密码子 91 突变的研究中选择了 gyrA 基因型结果为不确定、WT 和突变的 DNA 提取物。我们使用最近获得 CE 标记的 ResistancePlus GC（beta）检测（澳大利亚悉尼 SpeeDx）重新检测了这些标本。
结果
在高分辨率熔解分析中，86 份 gyrA 基因型不确定的标本中，ResistancePlus GC（beta）检测（SpeeDx）鉴定出 30 份（35%）WT、22 份（26%）突变和 34 份（40%）不确定的 gyrA 基因型。
结论
与之前的 gyrA 基因分型高分辨率熔解分析相比，ResistancePlus GC（beta）检测在淋病奈瑟菌 gyrA 基因型确定方面显示出了更好的效果。
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