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Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
Occurrence of NDM-1, VIM-1, and OXA-10 Co-Producing Providencia rettgeri Clinical Isolate in China
Siquan Shen1,2, Xiangning Huang3, Qingyu Shi1,2
1Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai, China.
Abstract:
Providencia rettgeri is a nosocomial pathogen associated with urinary tract infections related to hospital-acquired Infections. In recent years, P. rettgeri clinical strains producing New Delhi Metallo-β-lactamase (NDM) and other β-lactamase which reduce the efficiency of antimicrobial therapy have been reported. However, there are few reports of P. rettgeri co-producing two metallo-β-lactamases in one isolate. Here, we first reported a P. rettgeri strain (P138) co-harboring blaNDM-1, blaVIM-1, and blaOXA-10. The specie were identified using MALDI-TOF MS. The results of antimicrobial susceptibility testing by broth microdilution method indicated that P. rettgeri P138 was resistant to meropenem (MIC = 64μg/ml), imipenem (MIC = 64μg/ml), and aztreonam (MIC = 32μg/ml). Conjugation experiments revealed that the blaNDM-1-carrying plasmid was transferrable. The carbapenemase genes were detected using PCR and confirmed by PCR-based sequencing. The complete genomic sequence of the P. rettgeri was identified using Illumina (Illumina, San Diego, CA, USA) short-read sequencing (150bp paired-end reads), and many common resistance genes had been identified, including blaNDM-1, blaVIM-1, blaOXA-10, aac(6')-Il, aadA5, ant(2'')-Ia, aadA1, aac(6')-Ib3, aadA1, aph(3')-Ia, aac(6')-Ib-cr, qnrD1, qnrA1, and catA2. The blaNDM-1 gene was characterized by the following structure: IS110-TnpA-IntI1-aadB-IS91-GroEL-GroES-DsbD-PAI-ble-blaNDM-1-IS91-QnrS1-IS110. Blast comparison revealed that the blaNDM-1 gene structure shared >99% similarity with plasmid p5_SCLZS62 (99% nucleotide identity and query coverage). In summary, we isolated a P. rettgeri strain coproducing blaNDM-1, blaVIM-1, and blaOXA-10. To the best of our acknowledge, this was first reported in the world. The occurrence of the strain needs to be closely monitored.
Insights
This study reports the first isolation of a Providencia rettgeri strain co-producing three key beta-lactamase genes: blaNDM-1, blaVIM-1, and blaOXA-10. This finding highlights a significant challenge in treating hospital-acquired infections with multi-drug resistant bacteria.
Area of Science:
- Microbiology
- Infectious Diseases
- Genomics
Background:
- Providencia rettgeri is a significant nosocomial pathogen.
- Emergence of beta-lactamase-producing strains complicates antimicrobial therapy.
- Co-production of multiple metallo-beta-lactamases in P. rettgeri is rarely reported.
Purpose of the Study:
- To report the first isolation of a P. rettgeri strain co-harboring blaNDM-1, blaVIM-1, and blaOXA-10.
- To characterize the antimicrobial resistance profile and genetic elements associated with this multi-drug resistant isolate.
Main Methods:
- Species identification using MALDI-TOF MS.
- Antimicrobial susceptibility testing via broth microdilution.
- PCR, sequencing, and whole-genome sequencing for gene identification and characterization.
- Conjugation experiments for plasmid transferability.
Main Results:
- A P. rettgeri strain (P138) was identified, co-harboring blaNDM-1, blaVIM-1, and blaOXA-10.
- The isolate exhibited resistance to meropenem, imipenem, and aztreonam.
- The blaNDM-1 gene was located on a transferable plasmid.
- Whole-genome sequencing identified numerous resistance genes, including blaNDM-1, blaVIM-1, blaOXA-10, and qnr genes.
Conclusions:
- This is the first report globally of a P. rettgeri strain co-producing NDM-1, VIM-1, and OXA-10 beta-lactamases.
- The presence of multiple carbapenemase genes confers significant antimicrobial resistance.
- Close monitoring of such multi-drug resistant strains is crucial for infection control.
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