Capture ELISA for KPC Detection in Gram-Negative Bacilli: Development and Standardisation

The detection of KPC-type carbapenemases is necessary for guiding appropriate antibiotic therapy and the implementation of antimicrobial stewardship and infection control measures. Currently, few tests are capable of differentiating carbapenemase types, restricting the lab reports to their presence or not. The aim of this work was to raise antibodies and develop an ELISA test to detect KPC-2 and its D179 mutants. The ELISA-KPC test was designed using rabbit and mouse polyclonal antibodies. Four different protocols were tested to select the bacterial inoculum with the highest sensitivity and specificity rates. The standardisation procedure was performed using 109 previously characterised clinical isolates, showing 100% of sensitivity and 89% of specificity. The ELISA-KPC detected all isolates producing carbapenemases, including KPC variants displaying the ESBL phenotype such as KPC-33 and -66.

Automated summary

Detection of KPC-type carbapenemases is important for guiding antibiotic therapy and infection control measures. This study aimed to develop an ELISA test to specifically detect KPC-2 and its D179 mutants. The ELISA-KPC test showed high sensitivity and specificity, successfully detecting all isolates producing carbapenemases, including those with ESBL phenotype.