The Broad Institute of MIT and Harvard, Boston, Massachusetts, USA.
Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts, USA.
J Clin Microbiol. 2024 Nov 13;62(11):e0099724. doi: 10.1128/jcm.00997-24. Epub 2024 Oct 21.
Antimicrobial resistance is a growing health threat, but standard methods for determining antibiotic susceptibility are slow and can delay optimal treatment, which is especially consequential in severe infections such as bacteremia. Novel approaches for rapid susceptibility profiling have emerged that characterize either bacterial response to antibiotics (phenotype) or detect specific resistance genes (genotype). entypic and enotypic through NA detection (GoPhAST-R) is a novel assay, performed directly on positive blood cultures, that integrates rapid transcriptional response profiling with the detection of key resistance gene transcripts, thereby providing simultaneous data on both phenotype and genotype. Here, we performed the first clinical pilot of GoPhAST-R on 42 positive blood cultures: 26 growing , 15 growing , and 1 with both. An aliquot of each positive blood culture was exposed to nine different antibiotics, lysed, and underwent rapid transcriptional profiling on the NanoString platform; results were analyzed using an in-house susceptibility classification algorithm. GoPhAST-R achieved 95% overall agreement with standard antimicrobial susceptibility testing methods, with the highest agreement for beta-lactams (98%) and the lowest for fluoroquinolones (88%). Epidemic resistance genes including the extended spectrum beta-lactamase and the carbapenemase were also detected within the population. This study demonstrates the clinical feasibility of using transcriptional response profiling for rapid resistance determination, although further validation with larger and more diverse bacterial populations will be essential in future work. GoPhAST-R represents a promising new approach for rapid and comprehensive antibiotic susceptibility testing in clinical settings.IMPORTANCEExposure to antibiotics causes differential transcriptional signatures in susceptible vs resistant bacteria. These differences can be leveraged to rapidly predict resistance profiles of and in clinically positive blood cultures.
抗生素耐药性是一个日益严重的健康威胁，但确定抗生素敏感性的标准方法耗时且可能延误最佳治疗，这在严重感染（如菌血症）中尤为重要。已经出现了一些新的快速药敏分析方法，这些方法要么描述细菌对抗生素的反应（表型），要么检测特定的耐药基因（基因型）。通过 NA 检测进行的 entypic 和 enotypic（GoPhAST-R）是一种新的检测方法，直接在阳性血培养物上进行，它将快速转录反应分析与关键耐药基因转录本的检测相结合，从而同时提供表型和基因型的数据。在这里，我们对 42 个阳性血培养物进行了首次临床试点 GoPhAST-R 检测：26 个生长菌，15 个非生长菌，1 个同时存在生长菌和非生长菌。每个阳性血培养物的一部分被暴露于九种不同的抗生素中，裂解后在 NanoString 平台上进行快速转录谱分析；使用内部药敏分类算法分析结果。GoPhAST-R 与标准抗菌药敏检测方法的总体一致性达到 95%，其中β-内酰胺类药物的一致性最高（98%），氟喹诺酮类药物的一致性最低（88%）。流行的耐药基因，包括扩展谱β-内酰胺酶和碳青霉烯酶，也在该群体中被检测到。这项研究证明了使用转录反应谱进行快速耐药鉴定的临床可行性，尽管在未来的工作中，还需要用更大和更多样化的细菌群体进行进一步验证。GoPhAST-R 代表了一种在临床环境中快速全面进行抗生素药敏测试的有前途的新方法。
重要性抗生素暴露会导致敏感菌和耐药菌的转录特征存在差异。这些差异可以被利用来快速预测临床阳性血培养物中 的耐药谱。
