Comparison between polymerase chain reaction-based and checkerboard DNA hybridization techniques for microbial assessment of subgingival plaque samples.
OBJECTIVE
To compare polymerase chain reaction (PCR) with subsequent reverse hybridization (micro-IDent test) and checkerboard DNA-DNA hybridization for the identification of 13 bacterial species in subgingival plaque samples.


METHODS
Subgingival plaque samples were taken using paper points and curettes from two sites each with pocket depth <4, 4-6 and >6 mm at baseline and 3 months in 25 periodontitis subjects and two sites in 25 periodontally healthy subjects. Samples were analysed for their content of 13 bacterial species using both assays. Similarities for each species between techniques were determined using regression analysis. Differences between health and periodontitis were determined using the Mann-Whitney test.


RESULTS
Three hundred and fifty samples were evaluated using both techniques. Regression analysis indicated that 10/13 test species showed significant positive correlations between the counts determined by checkerboard analysis and levels determined by the PCR-based test after adjusting for 13 comparisons. The highest rank correlations of 0.58, 0.49 and 0.46 were seen for Treponema denticola, Fusobacterium nucleatum and Eubacterium nodatum, respectively (p<0.0001). Both tests could distinguish samples from healthy and periodontitis subjects.


CONCLUSIONS
Detection patterns of 10/13 test species in subgingival plaque samples from periodontitis and healthy subjects were similar using the two molecular techniques.
Haffajee
Anne D
AD
Department of Periodontology, The Forsyth Institute, Boston, MA 02115, USA. ahaffajee@forsyth.org
Yaskell
Tina
T
Torresyap
Gay
G
Teles
Ricardo
R
Socransky
Sigmund S
SS
D016976:Aggregatibacter actinomycetemcomitans / Q000302:isolation & purification; D001419:Bacteria / Q000145:classification* / Q000302:isolation & purification; D001439:Bacteroides / Q000302:isolation & purification; D044883:Campylobacter rectus / Q000302:isolation & purification; D002206:Capnocytophaga / Q000145:classification / Q000302:isolation & purification; D055113:Chronic Periodontitis / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis*; D003773:Dental Plaque / Q000382:microbiology*; D016124:Eikenella corrodens / Q000302:isolation & purification; D005051:Eubacterium / Q000302:isolation & purification; D016967:Fusobacterium nucleatum / Q000302:isolation & purification; D005884:Gingival Hemorrhage / Q000382:microbiology; D005891:Gingivitis / Q000382:microbiology; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010462:Peptostreptococcus / Q000302:isolation & purification; D017622:Periodontal Attachment Loss / Q000382:microbiology; D010514:Periodontal Pocket / Q000382:microbiology; D010519:Periodontium / Q000382:microbiology; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis / Q000302:isolation & purification; D018713:Prevotella intermedia / Q000302:isolation & purification; D048373:Treponema denticola / Q000302:isolation & purification; D055815:Young Adult
D004269:DNA, Bacterial