AUTHOR=Liao Wenjian , Long Dan , Huang Qisen , Wei Dandan , Liu Xiaobing , Wan Lagen , Feng Yuling , Zhang Wei , Liu Yang 

TITLE=Rapid Detection to Differentiate Hypervirulent Klebsiella pneumoniae (hvKp) From Classical K. pneumoniae by Identifying peg-344 With Loop-Mediated Isothermal Amplication (LAMP)

JOURNAL=Frontiers in Microbiology

VOLUME=11

YEAR=2020

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.01189

DOI=10.3389/fmicb.2020.01189

ISSN=1664-302X

ABSTRACT=<sec><title>Objectives</title><p>To establish a rapid molecular diagnostics of hvKp using the <italic>peg-344</italic> loop-mediated isothermal amplification technique (LAMP).</p></sec><sec><title>Methods</title><p>In all, 28 <italic>K. pneumoniae</italic> strains isolated from the blood of patients were used for the <italic>peg-344</italic> LAMP. <italic>K. pneumoniae</italic> NTUH-K2044 and <italic>K. pneumoniae</italic> ATCC700603 were used as positive control and negative control, respectively. For comparison, all the results were detected in a polymerase chain reaction (PCR), which was considered the gold standard for the detection of the gene. Mouse lethality assay, and Serum killing assay were also used to determine the virulence phenotype of <italic>K. pneumoniae</italic>.</p></sec><sec><title>Results</title><p>We determined the specificity and sensitivity of the primers for <italic>peg-344</italic> detection in the LAMP reactions. This LAMP assay was able to specifically differentiate hvKp from classical <italic>K. pneumoniae</italic> (cKp) at 65°C, which was 100-fold more sensitive than a PCR assay for <italic>peg-344</italic> detection. The virulence phenotype of <italic>K. pneumoniae</italic> detected by LAMP was as precise as by Mouse lethality assay and Serum killing assay.</p></sec><sec><title>Conclusion</title><p>The LAMP assay is easy to perform and rapid. Therefore, it can be routinely applied to differentiate hvKp from cKp in the clinical laboratory.</p></sec>