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Identification of hypervirulent Klebsiella pneumoniae isolates using the string test in combination with Galleria mellonella infectivity

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Abstract

Distinguishing between hypervirulent Klebsiella pneumoniae (hvKp) and classical Klebsiella pneumoniae (cKp) is a challenge to clinical laboratories. The aim of this study was to determine the practicability of combining the G. mellonella killing assay with a string test to differentiate hvKp from cKp. One hundred and three clinical K. pneumoniae isolates were collected. PCR amplification and wzi sequencing were used to determine the capsular serotype. Virulence genes allS, iro, iuc, and rmpA2, used frequently to identify hvKp, were detected by PCR. The virulence of K. pneumoniae isolates was evaluated using the following assays in parallel: molecular markers detection, G. mellonella killing assay alone, G. mellonella killing assay combined with the string test, and mouse infection. The results showed that the sensitivity, specificity, positive predictive value, and negative predictive value of combining the G. mellonella killing assay with a string test were 95.56%, 94.83%, 93.48%, and 96.49%, respectively, compared with mouse infection used as a positive reference. These values were significantly greater than those obtained using the G. mellonella killing assay only. The sensitivity, specificity, positive predictive value, and negative predictive value of allS, iro, iuc, and rmpA2 were greater than 77.78%, but less than combining the G. mellonella killing assay and string test. G. mellonella killing assay used in conjugation with the string test is a relatively simple and accurate method to assess K. pneumoniae virulence and differentiate between hvKp and cKp.

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Acknowledgments

Dr. Stephen H. Gregory (Providence, Rhode Island, USA) helped write and edit this manuscript.

Funding

This study was supported by grants from the Technology Commission of Shanghai Municipality (17411961700), Shanghai Municipal Health Commission (201640130).

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Authors and Affiliations

  1. Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, China

    Gang Li, Yu Tang, Xiaofei Jiang & Yuan Lu

  2. Research Center for Clinical Medicine, Jinshan Hospital, Shanghai Medical College, Fudan University, Shanghai, China

    Jimin Shi

  3. Department of Laboratory Medicine, Jinshan Hospital, Shanghai Medical College, Fudan University, Shanghai, China

    Yan Zhao

  4. State Key Laboratory for Microbial Metabolism and School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai, China

    Yingzhou Xie

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  1. Gang Li
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  2. Jimin Shi
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  5. Yu Tang
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Contributions

GL, XFJ, and YL conceived the study. GL, JMS, YZ, YT, YZ, and YZX performed the experiments and analyzed the data. XFJ and YL supervised the study.

Corresponding authors

Correspondence to Xiaofei Jiang or Yuan Lu.

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Ethics statement

Animal studies were performed in accordance with “Guide for the Care and Use of Laboratory Animals” (8th Ed.) and the protocol approved by Shanghai Public Health Clinical Center’s Animal Care and Use Committee.

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The authors declare that they have no conflicts of interest to declare.

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Li, G., Shi, J., Zhao, Y. et al. Identification of hypervirulent Klebsiella pneumoniae isolates using the string test in combination with Galleria mellonella infectivity. Eur J Clin Microbiol Infect Dis 39, 1673–1679 (2020). https://doi.org/10.1007/s10096-020-03890-z

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  • DOI: https://doi.org/10.1007/s10096-020-03890-z

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