Abstract

A rapid and accurate detection of carbapenemase-producing Gram-negative bacteria (CPGNB) has an immediate demand in the clinic. Here, we developed and validated a method for rapid detection of CPGNB using Blue-Carba combined with deep learning (designated as AI-Blue-Carba). The optimum bacterial suspension concentration and detection wavelength were determined using a Multimode Plate Reader and integrated with deep learning modeling. We examined 160 carbapenemase-producing and non-carbapenemase-producing bacteria using the Blue-Carba test and a series of time and optical density values were obtained to build and validate the machine models. Subsequently, a simplified model was re-evaluated by descending the dataset from 13 time points to 2 time points. The best suitable bacterial concentration was determined to be 1.5 optical density (OD) and the optimum detection wavelength for AI-Blue-Carba was set as 615 nm. Among the 2 models (LRM and LSTM), the LSTM model generated the higher ROC-AUC value. Moreover, the simplified LSTM model trained by short time points (0–15 min) did not impair the accuracy of LSTM model. Compared with the traditional Blue-Carba, the AI-Blue-Carba method has a sensitivity of 95.3% and a specificity of 95.7% at 15 min, which is a rapid and accurate method to detect CPGNB.

Highlights

  • Antimicrobial resistance (AMR) poses a serious global threat to human, animal, and environment health, as multidrug resistant bacteria continue to emerge and spread worldwide

  • The scanning results of positive and negative carbapenemase producers indicated a maximum absorption peak at 615 nm and this was chosen as the detection wavelength in following experiments (Supplementary Figure S1A)

  • We examined differing bacterial concentrations and diluents at these wavelengths and found that the optical density (OD) values varied

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Summary

Introduction

Antimicrobial resistance (AMR) poses a serious global threat to human, animal, and environment health, as multidrug resistant bacteria continue to emerge and spread worldwide. Carbapenems are one of the last-resort antibiotics to treat infections caused by multidrugresistant Gram-negative pathogens. Rapid Carbapenemase Producer Detection Assay primarily due to the production of various carbapenemases, which leaves the clinicians with limited therapeutic options. Carbapenemase-producers showed broad spectrum enzyme activity for various β-lactam substrates, and were associated with resistance to other antibiotic classes, and demonstrated rapid transmission in healthcare facilities, animals and the environments (Codjoe and Donkor, 2017). It is of paramount importance to develop reliable methods for rapid detection and characterization of carbapenemase-producers

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