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Objectives  To establish a rapid molecular diagnostics of hvKp using the  peg-344  loop-mediated isothermal amplification technique (LAMP).    Methods  In all, 28  K. pneumoniae  strains isolated from the blood of patients were used for the  peg-344  LAMP.  K. pneumoniae  NTUH-K2044 and  K. pneumoniae  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  K. pneumoniae .    Results  We determined the specificity and sensitivity of the primers for  peg-344  detection in the LAMP reactions. This LAMP assay was able to specifically differentiate hvKp from classical  K. pneumoniae  (cKp) at 65°C, which was 100-fold more sensitive than a PCR assay for  peg-344  detection. The virulence phenotype of  K. pneumoniae  detected by LAMP was as precise as by Mouse lethality assay and Serum killing assay.    Conclusion  The LAMP assay is easy to perform and rapid. Therefore, it can be routinely applied to differentiate hvKp from cKp in the clinical laboratory.

frontiers in microbiology

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