The effect of antibiotics and endodontic antimicrobials on the polymerase chain reaction.
The effectiveness of endodontic antimicrobial treatment could be determined using sensitive molecular methods. The purpose of this study was to determine if antibiotics or endodontic reagents interfere with the ability of PCR to detect Enterococcus faecalis in vitro. Amoxicillin (25 mg/ml), clindamycin (15 mg/ml), tetracycline (25 mg/ml), doxycycline (10 mg/ml), calcium hydroxide, 1% buffered sodium hypochlorite (NaOCl1), 3% and 6% unbuffered NaOCl (NaOCl3 and NaOCl6), 2% chlorhexidine (CHX), 5% tincture iodine (TI), 2% iodine potassium iodide (IKI), chloroform (CF), 70% ethyl alcohol, 5% sodium thiosulphate, 5% citric acid or saline were added to 10 or 10 cells/ml E. faecalis ATCC 19433 for 1 h (1 wk for Ca(OH)2). Using PCR, all specimens were positive except for NaOCl3 and NaOCl6. PCR with Ca(OH)2 was positive with 10 cells/ml but negative with 10 cells/ml. The following reagents yielded negative culturing results: all antibiotics, Ca(OH)2, CHX, IKI, TI, NaOCl3, NaOCl6, and CF. BacLight nuclear staining revealed the presence of viable cells in all PCR positive, culture negative combinations, except for those with CF. Therefore, in the presence of threshold values of bacterial concentrations, all reagents tested except for NaOCl3 and NaOCl6 do not interfere with the detection of E. faecalis using PCR.
Fouad
Ashraf F
AF
Department of Endodontology, School of Dental Medicine, University of Connecticut Health Center, Farmington CT, USA. aff001@dental.umaryland.edu
Barry
Jody
J
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D013293:Enterococcus faecalis / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000187:drug effects*; D011671:Pulpitis / Q000188:drug therapy / Q000382:microbiology; D012756:Sheep
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial