A proof-of-concept in vitro study to evaluate bacterial fluid path

transfer prevention in a newly developed peritoneal dialysis connector

Yekinni I , Viker T , Hunter R , Erdman A
1 2 3 1
1
Earl E. Bakken Medical Devices Center, University of Minnesota, Minneapolis, Minnesota, United States.

2
Cerovations, LLC, Saint Paul, Minnesota, United States.

3
Department of Microbiology & Immunology, University of Minnesota Medical School, Minneapolis, Minnesota, United States

Introduction/Objectives Results Discussion

More than 50% of peritoneal dialysis (PD) associated peritonitis episodes are from Microbial culture plates derived from the newly developed connector fluid path showed
bacteria colonizing the skin or mucous membrane and touch contamination with no growth in the 5 non-control tests (Group 2 - 6) irrespective of the culture medium.

patient handling of PD devices contributes to over 40% of episodes [1,2]. The major

COLONY FORMING UNITS (CFU)/mL

contamination prevention strategy utilized by PD programs today is training. PD 1 New connector 7 In comparison, microbial culture plates derived from the standard of care fluid path
1.00E+08 Sterile Contaminated
Nurses help patients master sterile exchange techniques and advise them to report Control 2 3 4 5 6 Control showed growth in 5 of 5 non-control on Trypticase soy agar (Figure 2A, Figure 2B),

technique breaches so they can receive prophylactic antibiotics. However, over 50% of 4 of 5 non-control tests on Pseudomonas Isolation Agar (Figure 3A, Figure 3B),

1.00E+06
patients become less adherent to training after 6 months of commencing PD [3].
and 2 of 5 non-control tests on Mannitol salts agar (Figure 4A, Figure 4B).

In this study, we compared the incidence of bacterial transfer to fluid path between a 1.00E+04
newly developed connector and the standard of care after simulated contamination.
1.00E+02
Conclusion
Materials & Methods Results show that the new PD connector may reduce risk of fluid path contamination
1.00E+00 even in the presence of heavy contamination.
1 2 3 4 5 6 7
Materials included prototypes of the newly developed connector; samples of a Sterile Contaminated
Control Control
commonly used PD connector system (standard of care); PD solution; disconnect Y- GROUP NUMBER
Sets; 100mL effluent sample bags; Staphylococcus epidermidis ATCC1228 and
Pseudomonas aeruginosa ATCC27853, representing the most common Gram positive New connector Standard of Care
1
Sterile
2 3 4 5 6 7
Contaminated Acknowledgement
Control Standard of Care Control
and Gram negative bacteria observed in peritonitis [4] were used as test organisms.

The team obtained approval from the University of Minnesota Institutional Review Research reported in this publication was supported by the National Institute of
Figure 2A. Growth on Trypticase soy agar Figure 2B. Microbial culture plates, Trypticase soy agar
Board and the Biosafety Committee.
Diabetes and Digestive and Kidney Diseases of the National Institute of Health under

award number DK126586.

To simulate contamination, 40mL of a standardized inoculum [1x108 colony-forming
units (CFU) per millilitre] was sprayed while a test participant connected a source of PD
COLONY FORMING UNITS (CFU)/mL

solution and a catheter extension set with gloved hands. To simulate patient 1.00E+08
1 New connector 7
Conflict of Interest
peritoneum and effluent, the catheter extension sets were pre-attached to disconnect Sterile Contaminated
Control 2 3 4 5 6 Control
Y-Sets and after connection attempts, test samples were collected in 100mL effluent Yekinni I is the inventor of the newly developed connector. Viker T is an employee of
1.00E+06
sample bags. The experimental setup was within a laminar flow biosafety cabinet in a Cerovations LLC, a company that has licensed the new connector from the University
biosafety level (BSL) 2 laboratory space. of Minnesota.
1.00E+04

1.00E+02 References
[1] Muthucumarana, K. et al. (2016) ‘The Relationship Between Presentation and the Time of Initial
1.00E+00
1 2 3 4 5 6 7 Administration of Antibiotics With Outcomes of Peritonitis in Peritoneal Dialysis Patients: The PROMPT Study’,
Sterile Contaminated Kidney International Reports. Elsevier Inc, 1(2), pp. 65–72. doi: 10.1016/j.ekir.2016.05.003.

Control Control

GROUP NUMBER
[2] Firanek, C. and Guest, S. (2011) ‘Hand hygiene in peritoneal dialysis’, Peritoneal Dialysis International, pp.
399–408. doi: 10.3747/pdi.2010.00239.

1 2 3 4 5 6 7

New connector Standard of Care Sterile Contaminated

Control Standard of Care Control [3] Segal, J. H. and Messana, J. M. (2013) ‘Prevention of peritonitis in peritoneal dialysis’, Seminars in Dialysis,
26(4), pp. 494–502. doi: 10.1111/sdi.12114.

Figure 3A. Growth on Pseudomonas Isolation Agar Figure 3B. Microbial culture plates, Pseudomonas Isolation Agar [4] Di Bonaventura, G. et al. (2012) ‘In vitro microbiology studies on a new peritoneal dialysis connector’,
Peritoneal Dialysis International, 32(5), pp. 552–557. doi: 10.3747/pdi.2011.00089.
COLONY FORMING UNITS (CFU)/mL

1.00E+08 New connector 7

1
Sterile Contaminated
Control 2 3 4 5 6 Control
1.00E+06

1.00E+04
Figure 1. Experimental setup with newly developed connector prototype
1.00E+02
Test sequence for both newly developed connector and standard of care included 7
samples obtained as follows: (I) a sterile control sample (Group 1) before exposure to
test organisms; (II) 5 test samples (Group 2 - 6) during connection attempts with 1.00E+00
1 2 3 4 5 6 7
simulated contamination; and (III) a contaminated control sample (Group 7) in which Sterile Contaminated
100mL of standardized inoculum in 2Liters of PD solution is allowed to flow through Control Control
GROUP NUMBER
the fluid path. Connection attempts were alternated between newly developed
connector and standard of care throughout the experiment.
1 2 3 4 5 6 7
New connector Standard of Care Sterile Contaminated

Control Standard of Care Control

The collected samples were maintained at body temperature (37° C) for 24 hours
before being inoculated on to microbial culture plates and media. Trypticase soy agar, Figure 4A. Growth on Mannitol salts agar Figure 4B. Microbial culture plates, Mannitol salts agar
Pseudomonas Isolation Agar and Mannitol salts agar were used.