Abstract:

Klebsiella pneumoniae which can cause diseases such as pneumonia was identified as a new foodborne
pathogenic bacterium in recent years. Using an ESE-Quant tube scanner, a new real-time fluorescent loop-mediated
isothermal amplification method for detecting K. pneumoniae was established with the primers designed based on the phoE
gene sequence published in GenBank. The results showed that this method had high specificity and sensitivity than PCR
method. Among 21 tested strains, 4 were identified as K. pneumoniae, and the other 17 were negative. The detection limit of
K. pneumoniae in artificially contaminated milk powder was 79 CFU/g, 100 times more sensitive than the conventional PCR
method. Moreover, the positive results could be directly identified immediately through the data and chart collected by the
ESE-Quant tube scanner so that real-time results could be provided.

Key words: Klebsiella pneumoniae, phoE gene, real-time fluorescence LAMP, detection, SYBR Green Ⅰ