Gonçalves Ana Beatriz, Alves Valquíria, Neves Isabel, Read Antónia, Pinheiro Natália, Henius Anna E, Hasman Henrik, Peixe Luísa, Novais Ângela
UCIBIO, i4HB, Faculdade de Farmácia da Universidade do Porto, Rua Jorge Viterbo Ferreira 228, Porto 4050-313, Portugal.
Clinical Microbiology Laboratory, Local Healthcare Unit, Matosinhos, Portugal.
J Antimicrob Chemother. 2025 Jul 1;80(7):2025-2031. doi: 10.1093/jac/dkaf170.
Expansion of carbapenemase-producing Klebsiella pneumoniae (CP-Kp) is driven by within-hospital transmission, requiring timely typing data for effective infection control.
We evaluated real-time performance and flexibility of our previously developed Fourier-transform infrared (FT-IR) spectroscopy workflow (spectra acquisition and analysis by machine-learning model).
All CP-Kp infection isolates (n = 136) identified at a northern Portuguese hospital (April 2022-March 2023) were tested from Columbia agar with 5% sheep blood, identified by FT-IR (KL-type/sublineage) and confirmed by reference methods (wzi sequencing, MLST and/or WGS).
FT-IR typing from Columbia agar with 5% sheep blood showed 73% sensitivity, 79% specificity and 74% accuracy. Our method correctly identified 94% of typeable isolates, 87% of which were communicated in <24 h. Non-typeable isolates belonged to new KL-types to the model (40%) or non-recognized KL-types (60%), most of which (66%) were correctly predicted when retested from Mueller-Hinton agar. Accuracy was then higher (88%) when results from both culture media were considered, and the model retrained to incorporate new sublineages. Three K. pneumoniae sublineages (ST147-KL64, ST15-KL19, ST268-KL20) were predominant and 86% of the isolates were correctly identified. During the study, an outbreak by ST268-KL20 in the neonatal ICU was quickly recognized, and solved in 23 days. Most isolates (98%) produced KPC-3.
We demonstrate that FT-IR spectroscopy meets high performance standards in real-time and adaptability to clonal dynamics, and we provide practical guidance for integrating FT-IR into daily microbiology practices. The unique time to response (same day as bacterial identification) enables early and effective infection control interventions.
产碳青霉烯酶肺炎克雷伯菌(CP-Kp)的传播是由医院内传播驱动的,需要及时的分型数据以进行有效的感染控制。
我们评估了我们之前开发的傅里叶变换红外(FT-IR)光谱工作流程(通过机器学习模型进行光谱采集和分析)的实时性能和灵活性。
对葡萄牙北部一家医院(2022年4月至2023年3月)鉴定出的所有CP-Kp感染分离株(n = 136),用含5%绵羊血的哥伦比亚琼脂进行检测,通过FT-IR进行分型(KL型/亚系),并通过参考方法(wzi测序、多位点序列分型和/或全基因组测序)进行确认。
用含5%绵羊血的哥伦比亚琼脂进行FT-IR分型显示敏感性为73%,特异性为79%,准确性为74%。我们的方法正确鉴定了94%的可分型分离株,其中87%在<24小时内得到通报。不可分型的分离株属于模型中的新KL型(40%)或未识别的KL型(60%),当从穆勒-欣顿琼脂重新检测时,其中大部分(66%)被正确预测。当考虑两种培养基的结果并对模型进行重新训练以纳入新的亚系时,准确性更高(88%)。三个肺炎克雷伯菌亚系(ST147-KL64、ST15-KL19、ST268-KL20)占主导地位,86%的分离株被正确鉴定。在研究期间,新生儿重症监护病房发生的ST268-KL20暴发很快被识别,并在23天内得到解决。大多数分离株(98%)产生KPC-3。
我们证明FT-IR光谱在实时性和对克隆动态的适应性方面符合高性能标准,并且我们为将FT-IR整合到日常微生物学实践中提供了实用指导。独特的响应时间(与细菌鉴定同一天)能够实现早期和有效的感染控制干预。