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Abstract: FR-OR009

Peritoneal Fluid Cell Free DNA Is a Versatile Analyte of Peritonitis

Session Information

Category: Dialysis

  • 703 Dialysis: Peritoneal Dialysis

Authors

  • Lee, John Richard, Weill Cornell Medicine, New York, New York, United States
  • Srivatana, Vesh, The Rogosin Institute, New York, New York, United States
  • Burnham, Philip, Cornell University, Ithaca, New York, United States
  • Renaghan, Amanda DeMauro, Weill Cornell Medicine, New York, New York, United States
  • Guo, Xunxi Susan, Weill Cornell Medicine, New York, New York, United States
  • Westblade, Lars, Weill Cornell Medicine, New York, New York, United States
  • Edusei, Emmanuel Y., Weill Cornell Medicine, New York, New York, United States
  • Silberzweig, Jeffrey I., The Rogosin Institute, New York, New York, United States
  • Albakry, Shady Y., Weill Cornell Medicine, New York, New York, United States
  • Magruder, Matthew, Weill Cornell Medicine, New York, New York, United States
  • Suthanthiran, Manikkam, Weill Cornell Medicine, New York, New York, United States
  • De Vlaminck, Iwijn, Cornell University, Ithaca, New York, United States
Background

Culture negative peritonitis is common in peritoneal dialysis (PD) patients, especially in the setting of recent antibiotic use. We investigated whether sequencing cell free DNA (cfDNA) extracted from peritoneal fluid supernatants can identify the suspected causative agent(s) in cases of peritonitis.

Methods

We recruited 7 PD subjects: 3 subjects had peritonitis and 4 subjects did not have evidence of peritonitis; and we collected 16 peritoneal fluid specimens. cfDNA was isolated using a Qiagen Circulating Nucleic Acid Kit and a single stranded library preparation was constructed for each specimen. The library was sequenced using an Illumina Next-Seq instrument (75 base pair by 75 base pair). Pathogen identification was performed using the bioinformatics program, GRAMMy.

Results

We obtained approximately 10 million cfDNA reads per specimen. Three PD subjects had an episode of culture confirmed peritonitis (Klebsiella pneumoniae, Staphylococcus aureus, and Staphylococcus epidermidis). cfDNA profiling identified the organisms in all 3 cases and importantly on subsequent days after antibiotic administration which had corresponding same day cultures that were negative. One of the subjects subsequently developed another episode of suspected peritonitis. The subject was treated for peritonitis with intraperitoneal antibiotics, but the peritoneal fluid cultures on admission were negative. The subject continued to have abdominal pain despite intraperitoneal antibiotics. The subject was suspected to have cholecystitis and culltures from a percutaneous cholecystotomy 4 days after admission grew Enterococcus faecium and Parabacteroides distasonis. cfDNA profiling of peritoneal fluid detected Enterococcus faecium and Parabacteroides distasonis on admission.

Conclusion

Our pilot data support the use of cfDNA profiling of peritoneal fluid to investigate peritonitis, especially in the setting of antibiotic use, and potentially as a method to investigate occult infections in abdominal organs.

Funding

  • Private Foundation Support