Fritzenwanker Moritz, Grabitz Marcel O, Negwer Vera, Schwengers Oliver, Arneth Borros, Chakraborty Trinad, Imirzalioglu Can, Wagenlehner Florian
Institute of Medical Microbiology, Justus Liebig University Giessen, Giessen, Germany.
Clinic for Urology, Pediatric Urology and Andrology, Justus-Liebig-University, Giessen, Germany.
Eur J Clin Microbiol Infect Dis. 2025 Mar;44(3):703-715. doi: 10.1007/s10096-024-05030-3. Epub 2025 Jan 7.
We designed and tested a point of care test panel to detect E.coli and antibiotic susceptibility in urine samples from patients at the point of care in the urological department. The aim of this approach is to facilitate choosing an appropriate antibiotic for urinary tract infections (UTI) at first presentation in the context of increasing antibiotic resistance in uropathogens worldwide.
We analyzed 162 E.coli isolates from samples from a university urological department to determine phenotypic and genotypic resistance data. With this data we created customized LAMP (loop-mediated isothermal amplification) panels for a commercial machine with which to detect and possibly quantify E.coli and six antibiotic resistance determinants. In a second step we tested these panel(s) for diagnostic accuracy on 1596 urine samples and compared with routine microbiological culture.
E.coli was detected with 95.4% sensitivity and 96.1% specificity. Dynamics of the LAMP amplification could be used to gauge bacterial loads in the samples. Antibiotic sensitivity was detected with good negative (sensitive) predictive values: ampicillin 92.8%, ampicillin/sulbactam 96.4%, cefuroxime 92.8%, cefotaxime 97.8%, trimethoprim/sulfamethoxazole 96.5%, ciprofloxacin 96.8%.
The LAMP panel provided E.coli detection and sensitivity information within one hour and thus could principally guide initial antibiotic therapy upon patients presenting with UTI. The panel helps to select initial adequate antibiotic therapy as well as providing diagnostic stewardship. Follow up investigations will expand the test system to other uropathogens.
我们设计并测试了一种即时检测试剂盒，用于在泌尿外科对患者的尿液样本进行即时检测，以检测大肠杆菌及抗生素敏感性。这种方法的目的是在全球范围内尿路病原体抗生素耐药性增加的背景下，便于在首次就诊时为尿路感染（UTI）选择合适的抗生素。
我们分析了来自一所大学泌尿外科样本中的162株大肠杆菌分离株，以确定表型和基因型耐药数据。利用这些数据，我们为一台商用仪器创建了定制的环介导等温扩增（LAMP）试剂盒，用于检测并可能定量大肠杆菌及六种抗生素耐药决定因素。第二步，我们在1596份尿液样本上测试了这些试剂盒的诊断准确性，并与常规微生物培养进行比较。
检测大肠杆菌的灵敏度为95.4%，特异性为96.1%。LAMP扩增的动力学可用于评估样本中的细菌载量。检测抗生素敏感性的阴性（敏感）预测值良好：氨苄西林为92.8%，氨苄西林/舒巴坦为96.4%，头孢呋辛为92.8%，头孢噻肟为97.8%，甲氧苄啶/磺胺甲恶唑为96.5%，环丙沙星为96.8%。
LAMP试剂盒可在一小时内提供大肠杆菌检测和敏感性信息，因此原则上可为患有UTI的患者指导初始抗生素治疗。该试剂盒有助于选择初始适当的抗生素治疗以及提供诊断管理。后续研究将把该检测系统扩展到其他尿路病原体。
