Development of a Simple Method to Detect the Carbapenemase-Producing Genes <i>bla</i>_NDM, <i>bla</i>_OXA-48-like, <i>bla</i>_IMP, <i>bla</i>_KPC, and <i>bla</i>_VIM Using a LAMP Method with Lateral Flow DNA Chromatography

Abstract

Infections by carbapenemase-producing Enterobacterales constitute a global public health threat. The rapid and efficient diagnosis of Enterobacterales infection is critical for prompt treatment and infection control, especially in hospital settings. We developed a novel loop-mediated isothermal amplification (LAMP) method combined with DNA chromatography to identify five major groups of carbapenemase-producing genes (blaNDM, blaOXA-48-like, blaIMP, blaKPC, and blaVIM). This method uses DNA–DNA hybridization-based detection in which LAMP products can be easily visualized as colored lines. No specific technical expertise, expensive equipment, or special facilities are required for this method, allowing its broad application. Here, 73 bacteria collections including strains with carbapenemase-producing genes were tested. Compared to sequencing results, LAMP DNA chromatography for five carbapenemase-producing genes had a sensitivity and specificity of 100% and >97%, respectively. This newly developed method can be a valuable rapid diagnostic test to guide appropriate treatments and infection control measures, especially in resource-limited settings.

Keywords

• loop-mediated isothermal amplification (LAMP)
• DNA chromatography
• carbapenem resistance
• carbapenemase-producing genes