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Rapid Pathogen Detection in Severe Infectious Keratitis: Prospective Evaluation of Combined Multiplex PCR Panels
Thomas Ferreira de Moura1,2, Sarah Gabison3, Yohan N'Guyen4,5
1Université de Reims Champagne-Ardenne; Service d'ophtalmologie, CHU de Reims, Bâtiment D, Avenue du Général Koenig, 51100, Reims, France. tferreira-de-moura@chu-reims.fr.
Introduction:
Rapid pathogen identification in severe infectious keratitis is critical for targeted therapy to prevent vision loss, but conventional methods are slow and only moderately sensitive. Our objective was to evaluate the diagnostic accuracy and turnaround time of combined FilmArray® Meningitis-Encephalitis (ME)/Blood Culture Identification (BCID) multiplex polymerase chain reaction (PCR) panels versus standard microbiology (culture + pathogen-specific PCR) in severe infectious keratitis.
Methods:
This was a prospective pilot diagnostic study at a French tertiary center (July 2023-April 2025) including 23 adults with severe microbial keratitis. Corneal samples were analyzed using both conventional microbiological testing (culture and pathogen-specific PCR) and rapid multiplex PCR with the FilmArray® ME and BCID panels. Primary outcomes were diagnostic agreement between the FilmArray® multiplex PCR system and conventional microbiological methods, measured by Cohen's κ coefficient. Turnaround times were compared using the Wilcoxon signed-rank test.
Results:
Among 23 adult patients (mean age 61.0 [SD 21.2] years, range 24-99; 13 female patients [56.5%]), conventional microbiology identified pathogens in 16 cases (69.5%) versus 15 (65.2%) with multiplex PCR. Overall diagnostic agreement was moderate (κ = 0.50; 95% CI, 0.25-0.76), with perfect concordance (12/12, 100%) for monomicrobial infections detected by both methods. Multiplex PCR significantly reduced mean time to identification (2 h [no variation]) versus conventional methods (102 [SD 42.5] h; P < .001). Both methods were negative in six patients (26.1%) with comparable clinical severity.
Conclusion:
Combined FilmArray® ME/BCID panels demonstrated complete concordance with standard microbiology for monomicrobial keratitis and reduced turnaround times by > 100 h. This strategy may accelerate targeted therapy, potentially improving visual outcomes.
Insights
Rapid multiplex PCR testing significantly cuts diagnostic time for severe infectious keratitis, offering faster targeted therapy. This approach shows high accuracy for monomicrobial infections, potentially improving patient vision outcomes.
Area of Science:
- Ophthalmology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Severe infectious keratitis requires rapid pathogen identification for effective treatment.
- Conventional diagnostic methods are often slow and lack sensitivity.
- Timely diagnosis is crucial to prevent vision loss.
Purpose of the Study:
- To evaluate the diagnostic accuracy of combined FilmArray® Meningitis-Encephalitis (ME)/Blood Culture Identification (BCID) multiplex PCR panels.
- To compare the turnaround time of multiplex PCR with standard microbiology for severe infectious keratitis.
- To assess the utility of rapid multiplex PCR in guiding targeted therapy.
Main Methods:
- Prospective pilot diagnostic study involving 23 adult patients with severe microbial keratitis.
- Corneal samples analyzed by conventional microbiology (culture, pathogen-specific PCR) and multiplex PCR (FilmArray® ME/BCID).
- Diagnostic agreement measured by Cohen's κ coefficient; turnaround times compared using Wilcoxon signed-rank test.
Main Results:
- Multiplex PCR identified pathogens in 65.2% of cases, compared to 69.5% with conventional methods.
- Overall diagnostic agreement was moderate (κ=0.50), with 100% concordance for monomicrobial infections.
- Multiplex PCR reduced mean identification time to 2 hours versus 102 hours for conventional methods (P<.001).
Conclusions:
- Combined FilmArray® ME/BCID panels offer rapid and accurate diagnosis for monomicrobial keratitis.
- This multiplex PCR strategy significantly decreases turnaround time, enabling faster targeted therapy.
- Accelerated diagnosis and treatment may lead to improved visual outcomes in patients with severe infectious keratitis.
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