6 Objectives Identify high risk group and individual Determine need for preventive measuresTo serve as an index of the success of the therapeutic measures.To motivate and to monitor the effectiveness of education programsManage the progress of restorative procedures.
7 Uses for Clinician Determine need for caries control measure Indicator of patient co-operationAid in timing of recall appointmentsGuide to insertion of expensive restorationsDetermine the prognosisPrecautionary signal to the orthodontist in placing bands
8 IDEAL REQUISITES Sound theoretical basis Maximum correlation with clinical status.Accurate with respect to duplication of results.SimpleQuick in performanceInexpensive , feasibleValidityReliabilitySensitivitySpecificity
10 1. LACTOBACILLUS COLONY COUNT TEST introduced by HADLEY in 1933PrincipleThis test measures the number of acidogenic and aciduric bacteria in patient’s saliva by counting the number of colonies on tomato peptone agar plates after inoculation with a sample of saliva.Cariology , 3rd Ed. - Earnest Newbrun
11 Procedurepatient chews a small piece of paraffinbefore breakfastThe saliva that accumulates in the 3 minute period is collected in a sterile containerSaliva collected is shaken by a machine for 2 minutes
12 0.4 ml of each sample is spread on the surface of an agar plate The saliva sample is diluted with sterile saline solution to 1:1000.4 ml of each sample is spread on the surface of an agar plateThe plates are incubated for 3 to 4 days at 37°CThe number of Lactobacillus colonies that develop are counted using Quebec counter.
13 Interpretation of LB colony count test No. of organismCFU/mlSymbolic designationDegree of caries activity+Little or NoSlight++1,000– 5000Moderate+++,000Marked++++> 10,000
14 Disadvantages Quick and easy but results are available in days. Counting colony tediouscomplex instrument, trained personnel requiredNot completely exclude the growth of other relatively aciduric organismsInaccurate for predicting the onset of caries
15 2. Colorimetric SNYDER TEST SNYDER in 1951PrincipleTest measures the ability of salivary microorganisms to form organic acid from a carbohydrate mediumBromocresol green indicator dye, changes colour from green to yellow at pH 5.4 to 3.8Essentials of Preventive and community Dentistry- Soben Peter
16 ProcedureSalivary sample is collected after the patient chews a small piece of paraffin before breakfast.0.2 cc of saliva is pipetted into the 10ml melted agar containing medium in a test tube at 50°C, allowed to solidify and then incubated at 37°C .The incubation period is upto 72 hours.
17 . The media contains : Bactopeptone Dextrose Sodium chloride Agar Bromocresol greenThe rate of colour change from green to yellow is indicative of the degree of caries activity..
19 Advantages :relatively simpleModerate costDisadvantages :Time consumingSometimes color change is not very clearLimited predictive value
20 3. THE SWAB TEST GRAINGER et al in 1965. Principle same as Synder test.Cariology , 3rd Ed. - Earnest Newbrun
21 ProcedureThe oral flora is sampled by swabbing the buccal surfaces of the teeth with a cotton applicator,subsequently incubated in the medium.The change in the pH following a 48 hour incubation is read on a pH meter or the colour change is read by the use of a colour comparator.
22 Interpretation :pH 4.1 and < 4.1 = Marked cariespH 4.2 to = ActivepH 4.5 to 4.6 = Slightly activepH 4.6 and over = Caries inactive
23 4. SALIVARY REDUCTASE TEST measures the activity of the reductase enzyme present in salivary bacteriaThis enzyme is involved in the reactions for the formation of products dangerous to the tooth surface.Rate at which the indicator molecule , diazoresorcinol, changes from blue to red to colorless
24 ProcedureSaliva is collected by chewing a special flavoured paraffin and expectorated directly into collection tubesThe sample is then mixed with the dye Diazo-resorcinolThe colour changes and the “Caries Conduciveness” reading is taken after 15 minutes
26 AdvantagesNo incubationQuick resultsDisadvantagesResults vary with time after food intake & after brushing
27 5.ALBAN TEST Simple modified Snyder test Main Features: softer medium that permits the diffusion of saliva and acids without melting the medium.simpler sampling procedure , patient expectorates directly into tubes containing the mediumEssentials of Preventive and community Dentistry- Soben Peter
28 following materials are required: Synder test agarA small scale to measure 60 grams.A 2 litter Pyrex glass to melt the medium.A funnel to dispense the medium into test tubes.mm test tubes with screw caps
29 Procedure :60gms of Snyder test agar is placed in 1 litre of water &suspension is brought to a boil over a low flame.When thoroughly melted the agar is distributedThe tubes are autoclaved for 15 minutes: allowed to cool and stored in a refrigerator.2 tubes are taken & the patient is asked to expectorate a small amount of saliva directly into the tubes.incubated at 370C for upto 4 days
30 observe for:Change of colour from bluish green (pH 5) to definite yellow (pH 4 or below).The depth in the medium to which the change has occurred.
31 Scale for Scoring:1. No colour change = ‘3/4’2. Beginning colour change = ‘+’(from top to medium down)3. One half colour change = ‘++’(from top down)4. Three fourths colour change = ‘+++’5. Total colour changes to yellow = ‘++++’
32 Advantages:Simplicity,Low cost,Diagnostic value when negative results are obtained.Motivational valueDisadvantages:More armamentariumSubjective evaluation
33 6. SALIVARY BUFFER CAPACITY ActionMeasures the no. of mls of acid required to lower the pH from 7 to 6.EquipmentpH meter0.05N lactic acidParaffinSterile glass jars containing small amount of oil.Cariology , 3rd Ed. - Earnest Newbrun
34 Procedure10ml of saliva collected under oil 1hr after eating5ml is taken into a beakerpH of saliva is adjusted to 7.0Lactic acid is added until pH of 6.0 is reached.The no. of ml of lactic acid needed to reduce pH is a measure of buffer capacity.Evaluation :Inverse relationship
35 7. STREPTOCOCCUS MUTANS LEVEL IN SALIVA measures the number of Streptococcus mutans colony forming units per unit volume of salivaIncubation is done on Mitis Salivarius Agarwith addition of sucrose (20%) & 0.2U bacitracin /ml which supresses the growth of most non-S.Mutans colonies
36 The samples of organisms is obtained by the use of tongue blades ProcedureThe samples of organisms is obtained by the use of tongue bladesSample then pressed against Mitis SalivariusBacitracin Agar in special Petri dishesincubated at 370C for 48 hrs in 95%& 5%CO2gas mixtureCariology , 3rd Ed. - Earnest Newbrun
37 Interpretation : Levels of Streptococcus Mutans > 10 5/ ml of saliva = unacceptable,
38 DisadvantagesDifficulty of distinguishing between a carrier state and cariogenic infection.S. Mutans may constitute less than 1% of total flora of plaque.S. Mutans tends to be located at specific sites only.Not convenient for chair side
39 8.Dip slide method Dentocult Larmas ,1975 S.Mutans levels in saliva ProcedureSaliva is collected and poured over the agar coated slide & allowed to dryBacitracin disks are placed in the middleCo2 tablet is inserted in the tube & then incubated for 48 hrs.
40 Colony density (small blue colonies) compared with model chart negligible<105> 106
41 9.Plaque / tooth pick method ActionSimple screening of diluted plaque sampleProcedurePlaque samples are collected from the gingival thirds of buccal tooth surfaces & placed in Ringer’s solutionSuspension is stretched across MSA platesIncubation at 370C for 72 hrsCariology , 3rd Ed. - Earnest Newbrun
42 10. S. mutans adherence method Saliva inoculated in MSB broth for 24 hrs at 37°CAfter growth, the supernatant removedCells adhering to the glass examined macroscopicallySCORENO OF COLONIES-No growth expressed+Few deposits from 1-10++Scattered deposits of smaller size+++Numerous minute deposits & more than 20 large deposits (indicates > 105 CFU/ml)wS. mutans adherence methodS. mutans dip-slide methodsS. mutans replicate techniqueSee for all three -newbrun42
43 10.Fosdick Calcium dissolution test PrincipleMeasures the mgs of powdered enamel dissolved in 4 hours, when patients saliva mixed with glucose and powdered enamel.Essentials of Preventive and community Dentistry- Soben Peter
44 AdvantagesCorrelation reported is goodDisadvantagesComplex equipmentExpensive, trained personnel.
45 11. DEWAR TEST Test similar to fosdick calcium dissolution test Except that in this test the pH of the mixture is measured instead of the amount of calcium dissolved by the acid.Essentials of Preventive and community Dentistry- Soben Peter
46 12. CARIOSTAT TEST Tsutomu Shimono 1974 This test has been reported to significantly correlate with counts of S.mutans in childrenuses a semisynthetic liquid containing sucrose, tryptose, a gram-negative bacteria growth inhibitor, and bromcresol green and bromcresol purple indicators.The test assesses acid production by cariogenic bacteria and thus indirectly assesses caries activityThe sensitivity and specificity of a colorimetric microbiological caries activity test (Cariostat) in preschool children
47 Interpretation of Cariostat test ScorepHColorCaries Activity7BlueCaries inactive15.5GreenSlight caries24.5Yellow - GreenModerate caries34.0YellowMarked caries
48 13.Oratest Rosenberg et al. in 1989 Principle The rate of oxygen depletion by micro organisms in expectorated milk samples
49 Under aerobic conditions the bacterial enzyme, aerobic dehydrogenase transfers electrons or protons to oxygen.Once oxygen gets utilized by the aerobic organisms and an anaerobic environment is attained, methylene blue acts as an electron acceptor and gets reduced to leucomethylene blueThe metabolic activity of the aerobic microorganism is reflected by the reduction of methylene blue to leucomethylene blue.Oratest: A Simple Chairside Aid for Caries Risk Assessment
50 AdvantagesSimple , inexpensive , less time consuming , reproducible and requiring no trained personnel.Used to monitor mouth rinse regime , denture hygine , gingival inflammation and plaque levels.Good educational and motivational tool for patient,school and community health programmes.Vehicle for test is nontoxic (milk)Anticipate the onset of cariesResults can provide dentist to command patient to reinforce motivation ,plaque control and behaviour
51 DisadvantagesIt does not identify a specific group of organism in a specific disease.It cannot accurately differentiate between the healthy state and between initial and progressive carious lesionLack of specificity since positive observation can be obtained in gingivitis and other oral ailments.
52 14. Salivary glucosyltransferase b Gtfs play a critical role in the pathogenesis of caries and furthermore, the enzymes are present in whole saliva.Therefore, hypothesized that the amount of Gtfs collectively or separately could be correlated with caries activity.chair side and provide results within minutesSalivary Glucosyltransferase B as a Possible Marker for Caries Activity-
53 2 ml of saliva was collected saliva was clarified by centrifugation,Clarified saliva was mixed in a 1:1 ratio with the coating buffer supplied in the kit and was coated onto 96-well platesAfter washing, antibodies to Gtf B, C and D were appliedintensity of the color in the wells, which correlated to the amount of purified Gtf present, was read in an ELISA reader
54 15. CariScreen Caries Susceptibility Test is done in one minute at chairsideATP bioluminescence technology, which is a test for light reaction, will measure on a scale from 0 to 9999.the levels of acid-producing bacteria by swabbing lingual surfaces of lower anterior teeth and then inserting the sample in the CariScreen meter to be read0 to 1500 is low risk and 1500 to 9999 is high riskThe ATP generates light once it reacts with luciferin, a light- emitting biological pigment/enzyme present in the CariScreen. The amount of light emission is in direct proportion to the amount of ATP and a well-known fact is that acid-producing bacteria contain 100 times more ATP than non-acid-producing strains
56 16. Carivissystem for assessment of caries activity relies on the unique properties of the Glowdent marker that detects actively occuring dental demineralisationThe amount of light emitted varies with the amount of minerals presentmore minerals equals more light.
57 Advantages of the Carivis System The Carivis caries assessment system marks a new approach in preventive dentistry. Its innovative design offers:Non-invasive, one-step methodCaries activity assessment is possible with a single visit to the dentistExtreme sensitivity and information on the extent of demineralisationAccess to all dental surfaces, including contacting surfaces of teeth and around restorationsEase of useSpeed; signal collection requires less than 10 seconds
60 References Cariology , 3rd Ed. - Earnest Newbrun Principles & practice of pedodontics – Arathi RaoDentistry for the child & adolescent – McDonaldTextbook of Pedodontics- Shobha TandonTextbook of Pediatric dentistry- S.G.DamleEssentials of Preventive and community Dentistry- Soben PeterChair side simple caries activity test: Ora test- Bhasin S, JISPPD 2006;24:2A Modified dip slide test for microbiological risk in caries assessment.B Thaweboon,srosori thaweboon etal,Southeast Asian J Trop Med Public health; :2:400-04Salivary Glucosyltransferase B as a Possible Marker for Caries Activity- A.M. Vacca Smith. Caries Res November; 41(6): 445–450Using a Caries Activity Test to Predict Caries Risk in Early Childhood. Michiko Nishimura, JADAOratest: A Simple Chairside Aid for Caries Risk Assessment -Arora Ruchia, Lahiri. Prathik. Kb, Masih Updes. Int. j of dental clinics 2009:1(1): 26-30The sensitivity and specificity of a colorimetric microbiological caries activity test (Cariostat) in preschool children. Pediatric Dentistry: July/Aug Volume1 6, 4