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Abstract Background Molecular technologies can rapidly identify pathogens, allowing for the correct antimicrobial agent to be used for initial treatment. Many laboratories have implemented molecular technologies to identify the causative agents of infections from various bodily sources. Screening for resistance genes to common antimicrobial agents provides high value to clinicians to help guide optimal antimicrobial therapy selection. This study compares the accuracy of laboratory performance for molecular multiplex platforms in the detection of Gram-negative resistance genes from American Proficiency Institutes Blood Pathogen Panel proficiency program in 2024. Methods American Proficiency Institute (API), an independent proficiency testing provider, reviewed the reporting of various molecular multiplex systems against simulated proficiency samples offered in 2024. Participants were instructed to only report resistance genes their systems were capable of testing for, and ensure they were appropriate for the organism recovered. Failure rates for participants were evaluated against an 80% participant consensus standard for detection of resistance genes. Results During the 2024 proficiency year, Gram-negative targets were included 7 times in 15 unique samples within the Blood Pathogen Panel program. Participants accurately identified all 7 Gram-negative organisms and reported out resistance genes with varying accuracy and appropriateness. Accuracy for correct reporting of resistance genes appropriate for the organism recovered ranged between 95.0%- 100%. Conversely, the number of laboratories reporting resistance genes that were inappropriate for the organism recovered ranged between 1.0% - 9.0%. Conclusion Based on the results reported by participants in API’s proficiency program, molecular multiplex platforms are accurately identifying Gram-negative resistance genes. However, many participants using both FDA approved and LDT systems reported results for resistance genes that were not applicable for the organism recovered. This inaccurate reporting could potentially lead to inappropriate antimicrobial therapies being prescribed.