Rapid Detection to Differentiate HypervirulentKlebsiella pneumoniae(hvKp) From ClassicalK. pneumoniaeby Identifyingpeg-344With Loop-Mediated Isothermal Amplication (LAMP)
Rapid Detection to Differentiate HypervirulentKlebsiella pneumoniae(hvKp) From ClassicalK. pneumoniaeby Identifyingpeg-344With Loop-Mediated Isothermal Amplication (LAMP)
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DOI：
10.3389/fmicb.2020.01189
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发表时间：
2020-06-04
影响因子：
5.2
通讯作者：
Liu, Yang
中科院分区：
文献类型：
--
作者：
Liao, Wenjian;Long, Dan;Liu, Yang
Objectives To establish a rapid molecular diagnostics of hvKp using thepeg-344loop-mediated isothermal amplification technique (LAMP). Methods In all, 28K. pneumoniaestrains isolated from the blood of patients were used for thepeg-344LAMP.K. pneumoniaeNTUH-K2044 andK. pneumoniaeATCC700603 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 ofK. pneumoniae. Results We determined the specificity and sensitivity of the primers forpeg-344detection in the LAMP reactions. This LAMP assay was able to specifically differentiate hvKp from classicalK. pneumoniae(cKp) at 65 degrees C, which was 100-fold more sensitive than a PCR assay forpeg-344detection. The virulence phenotype ofK. pneumoniaedetected 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.
