Sensitive and specific LAMP and multiplex qRT-PCR assays for detection of hypervirulent Klebsiella pneumoniae

Abstract

Hypervirulent Klebsiella pneumoniae (hvKP) pose significant challenges to clinical anti-infective treatment and has emerged as a major threat to global public health. In this study, we employed the loop-mediated isothermal amplification (LAMP) assays with OTG (orange to green) visual dye and multiplex quantitative real-time PCR (qRT-PCR) assay to rapidly detect hvKP. We determined the detection limits of the LAMP methods for K. pneumoniae, iroB, and iucA genes and the qRT-PCR assay for iroB, iucA, rmpA and rmpA2. Additionally, we assessed the sensitivity and specificity (both over 95 %) of the LAMP and qRT-PCR methods. The LAMP and qRT-PCR methods established in this study have been proven to be simpler and more reliable than traditional PCR methods, making them an improved choice for laboratory diagnosis.