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Focus on the diagnosis of pathogenic microorganisms in the reproductive tract and respiratory tract

Development of a cost-effective rapid detection of carbapenem-resistance Gram-negative bacteria laboratory workflow

2024-02-07 10:48:54

Spread of Carbapenemase-producing Enterobacterales (CPE) has become a major global public health threat and clinical concerns worldwide.

Background

Spread of Carbapenemase-producing Enterobacterales (CPE) has become a major global public health threat and clinical

concerns worldwide. Most carbapenemase genes are located on plasmids or integrons which is easily transferred horizontally to other isolates, leading to rapidly spread, especially in hospital settings. Current clinical laboratory methods for the detection of CPE might be labor-intensive, required overnight incubation, or special equipment, not cost-effective procedures.

Methods

In this study, we have developed and validated a cost-effective rapid detection of CPE laboratory workflow, especially for

isolates from severe infection patients. A set of pre-selected 26 enteric or non-fermentative strains with known CPE (10 KPC, 5 IMP, 7 NDM, 1 VIM, and 3 OXA-48) was used to develop and train the workflow. An additional 105 clinical isolates were used to validate the proposed workflow. Routine MALDI-tof was used to identify clinical isolates rapidly. Vitek2 Antibiotics Susceptibility Test (AST) cards were used for routine AST results. PCR and Next Generation DNA Sequencing (NGS) methods were used to confirm the antibiotics resistant genotypes of clinical CPE isolates. The proposed workflow, starting from the primary isolation of bacterial colonies, using MALDI-tof as rapid identification, and integrated with a cost-effective 15 minutes lateral flow CPE tests were evaluated.

Results

Of all clinical isolates evaluated, 41 isolates were confirmed CPE positive by PCR/NGS. As compared to PCR/NGS results, the proposed workflow achieved 97.1% of accuracy during the same day of primary isolation, whereas compared to routine next-day Vitek2 AST results achieved 95.2% of accuracy.

Conclusions

This proposed workflow can rapidly detect 22 KPC and 18 NDM isolates encountered with great confidence and cost-effectiveness one day earlier than conventional AST results. However, due to small sample sizes with VIM, IMP and OXA-48 encountered in this study, more isolates should be included for the workflow utility in these phenotypes.

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