Method for studying development of colonization and infection of dialysis catheters.
Microbial colonization and infection of dialysis catheters is the major cause of catheter failure. The present study aimed to develop a method to permit the study of microbial biofilm formation and antibiotic prophylaxis and therapy. Standard silicon rubber and silver-impregnated catheters were sectioned into 1 mm slices and placed into 1-cm x 2-cm culture slide chambers. Fresh clinical isolates were obtained from infected patients and suspended in concentrations of 10(3), 10(6), and 10(9) colony forming units (CFU) per milliliter in a variety of liquid suspending culture media, which included serum protein constituents. Antibiotics could be added to the suspending fluid to determine prophylactic activity, or at any time thereafter to determine therapeutic activity. The catheter sections were incubated with the microbial challenge for 6 hours, 12 hours, 24 hours, and 48 hours and then washed in flowing distilled water to remove unattached biofilm. They were then stained with acridine orange, which fluoresces microbial DNA, and were examined by confocal scanning laser microscopy. Biofilm formation, representing colonization and infection, were quantified by comparing the fluorescent pixel analysis of the uninoculated control with the challenged catheter. The method was reproducible and permitted quantitative analysis. Standard silicon rubber catheters demonstrated greater biofilm formation than silver-impregnated catheters. The method examines the factors involved in microbial colonization and infection, and in antibiotic prophylaxis and therapy.
Khaled
G H
GH
Department of Laboratory Medicine, Yale University School of Medicine, New Haven, Connecticut, USA.
Finkelstein
F O
FO
Carey
H B
HB
Wardlaw
S C
SC
Kliger
A S
AS
Edberg
S C
SC
D018441:Biofilms; D002408:Catheters, Indwelling / Q000382:microbiology*; D015169:Colony Count, Microbial; D016983:Enterococcus / Q000187:drug effects / Q000254:growth & development; D004866:Equipment Contamination; D066298:In Vitro Techniques; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000254:growth & development; D008826:Microbial Sensitivity Tests; D018613:Microscopy, Confocal; D010530:Peritoneal Dialysis / Q000295:instrumentation*; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000254:growth & development; D012826:Silicone Elastomers; D012834:Silver; D013211:Staphylococcus aureus / Q000187:drug effects / Q000254:growth & development; D013212:Staphylococcus epidermidis / Q000187:drug effects / Q000254:growth & development
D012826:Silicone Elastomers; D012834:Silver