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Volume 31, Number 1—January 2025
Research

Cefiderocol Resistance Conferred by Plasmid-Located Ferric Citrate Transport System in Klebsiella pneumoniae Carbapenemase–Producing K. pneumoniae

Riccardo Polani, Alice De Francesco, Dario Tomolillo, Irene Artuso, Michele Equestre, Rita Trirocco, Gabriele Arcari, Guido Antonelli, Laura Villa, Gianni Prosseda, Paolo Visca, and Alessandra CarattoliComments to Author 
Author affiliation: Sapienza University of Rome, Rome, Italy (R. Polani, A. De Francesco, D. Tomolillo, R. Trirocco, G. Arcari, G. Antonelli, G. Prosseda, A. Carattoli); University of Pavia, Pavia, ltaly (D. Tomolillo); Istituto Superiore di Sanità, Rome (I. Artuso, M. Equestre, L. Villa); University of Insubria, Varese, Italy (G. Arcari); Sapienza University Hospital “Policlinico Umberto I,” Rome (G. Antonelli); Roma Tre University, Rome (P. Visca)

Main Article

Figure 3

Expression analysis of siderophore receptor genes in the presence and absence of plasmidic fec gene cluster and ferric citrate inducer in study of cefiderocol resistance conferred by plasmid-located ferric citrate transport system in Klebsiella pneumoniae carbapenemase–producing K. pneumoniae. A) Transcription of the fecA genes in the DH5-α strain carrying R69c or R69c-FEC, determined by using primer pairs able to discern the chromosomal fecA allele from the K. pneumoniae fecA gene in the fecABCDE operon or a primer pair recognizing both chromosomal and plasmidic fecA alleles (Appendix 1). The relative quantitative analysis of the transcripts was based on the 2−ΔΔCT method (33). In both bar graphs, the relative values were calculated with respect to the transcript level observed in the R69c carrying strains and set to 1. B) Transcription of the siderophore receptor genes fiu, cirA, fepA, and fhuA in the R69c-FEC and R69c strains grown in the absence of ferric citrate or in the presence of 0.5 μM or 5.0 μM ferric citrate, relative to the R69c strain grown without ferric citrate, which is set to 1. The relative quantitative analysis of the transcripts was based on the 2−ΔΔCT method (33). Error bars represent SDs. Statistical significance was determined by using a paired 2-tailed Student t-test comparing the dataset obtained from the 2 strains grown under the same conditions. FEC, ferric citrate transport system.

Figure 3. Expression analysis of siderophore receptor genes in the presence and absence of plasmidic fec gene cluster and ferric citrate inducer in study of cefiderocol resistance conferred by plasmid-located ferric citrate transport system in Klebsiella pneumoniae carbapenemase–producing K. pneumoniae. A) Transcription of the fecA genes in the DH5-α strain carrying R69c or R69c-FEC, determined by using primer pairs able to discern the chromosomal fecA allele from the K. pneumoniae fecA gene in the fecABCDE operon or a primer pair recognizing both chromosomal and plasmidic fecA alleles (Appendix 1). The relative quantitative analysis of the transcripts was based on the 2−ΔΔCT method (33). In both bar graphs, the relative values were calculated with respect to the transcript level observed in the R69c carrying strains and set to 1. B) Transcription of the siderophore receptor genes fiu, cirA, fepA, and fhuA in the R69c-FEC and R69c strains grown in the absence of ferric citrate or in the presence of 0.5 μM or 5.0 μM ferric citrate, relative to the R69c strain grown without ferric citrate, which is set to 1. The relative quantitative analysis of the transcripts was based on the 2−ΔΔCT method (33). Error bars represent SDs. Statistical significance was determined by using a paired 2-tailed Student t-test comparing the dataset obtained from the 2 strains grown under the same conditions. FEC, ferric citrate transport system.

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