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. 2025 Mar;44(3):703-715.
doi: 10.1007/s10096-024-05030-3. Epub 2025 Jan 7.

Genome-based development and clinical evaluation of a customized LAMP panel to rapidly detect, quantify, and determine antibiotic sensitivity of Escherichia coli in native urine samples from urological patients

Affiliations

Genome-based development and clinical evaluation of a customized LAMP panel to rapidly detect, quantify, and determine antibiotic sensitivity of Escherichia coli in native urine samples from urological patients

Moritz Fritzenwanker et al. Eur J Clin Microbiol Infect Dis. 2025 Mar.

Abstract

Purpose: 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.

Methods: 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.

Results: 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%.

Conclusion: 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.

Keywords: Antibiotic susceptibility testing; Antimicrobial stewardship; Fast microbiology; Loop-mediated isothermal amplification; Point of care testing; Urinary tract infections.

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Conflict of interest statement

Declarations. Ethical approval: Our study has been approved by the ethics committee of the Faculty of Medicine, Justus-Liebig-University of Giessen, file number AZ 80/17. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Project strategy
Fig. 2
Fig. 2
Diagnostic Workflow: A Urine samples are screened for E. coli with LAMP reactions. Positive samples are then tested for specific resistance genes. B The same urine samples are analyzed in the microbiological laboratory using standard urine culture and phenotypic susceptibility testing
Fig. 3
Fig. 3
LAMP detection of E.coli in vitro, dilution series: E.coli was diluted and measured with LAMP. Higher E.coli concentrations mean that LAMP turns positive faster
Fig. 4
Fig. 4
LAMP amplification curves from point of care testing of urine samples Sensitivity Forecast by LAMP-Panel
Fig. 5
Fig. 5
AMS workflow evaluation

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