Enhancing Carbapenem Resistance Detection: LAMP Coupled with Melting Curve Analysis- for Rapid Molecular Diagnostics in Clinical Specimens

Abstract

Carbapenem-resistant Enterobacteriaceae have become a major clinical and public health challenge due to the need for rapid administration of effective antimicrobials and implementation of supplemental infection control practices. Identifying genes associated with carbapenem resistance is crucial for managing these cases. The timely initiation of effective antimicrobial therapy and infection control interventions to prevent spreading are critical. Therefore, rapid diagnostic tests for detecting CP-CRE are necessary. This study aimed to evaluate the use of LAMP and melting curve analysis in RT-PCR to identify carbapenem resistance genes (NDM and OXA) in clinical specimens. This study emphasizes the benefits of the LAMP strategy in comparison to traditional methods, demonstrating its effectiveness and practicality. LAMP has proven to be more sensitive than RT-PCR and conventional multiplex PCR, with a detection rate of 96.7% and 91.6% respectively. While LAMP has a slightly lower specificity rate of 88.7%, it remains a promising method for rapid and accurate identification. Furthermore, the study found that LAMP can detect bacterial DNA even in low quantities, with a limit of detection of 102 CFU/mL for both K. pneumoniae and E. coli. In settings where MDR bacteria are prevalent, this capability is crucial for early diagnosis and treatment. In conclusion, RT-PCR systems that combine melting curve analysis with LAMP offer promise for quick and precise identification of carbapenem resistance genes in clinical specimens. This approach can enhance the diagnosis and management of multidrug-resistant bacterial infections.