advanced diagnostic aids khushbu
TRANSCRIPT
![Page 1: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/1.jpg)
Advanced Diagnostic Aids
Dr. Khushbu mishraHKE’s S.N Dental college,
Gulbarga
![Page 2: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/2.jpg)
Contents
Introduction Limitations of conventional periodontal diagnosis Advances In Clinical Diagnosis Advances In Radiographic Assessment Advances Microbiologic Analysis Advances In Characterizing The Host Response
![Page 3: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/3.jpg)
Introduction
• Definition : Diagnosis is defined as identifying the disease from an evaluation of history, signs and symptoms, laboratory tests and procedures.
• Importance :
a. It identifies and indicates the nature of etiological factors
b. Indicates the nature of pathological processes
c. It is essential for treatment planning
![Page 4: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/4.jpg)
types of diagnosis
Provisional differential comprehensive
therapeutic
emergency
Diagnostic aids in periodontics conventional
advanced
![Page 5: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/5.jpg)
• Periodontal disease Gingival Inflammation
Periodontal Destruction
• Diagnosis :- clinical evaluation ( Gingivitis)
clinical evaluation
+ (periodontitis)
tissue destruction
![Page 6: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/6.jpg)
• Tissue destruction ( Periodontitis)
Loss of connective tissue
clinical radiographic
attatchment loss bone loss
Gives historical evidence of damage Identify and quantify current clinical signs of inflammation
![Page 7: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/7.jpg)
Limitations
• Does not provide cause of condition• Susceptibility of patient• Cannot reliably identify sites with ongoing periodontal
destruction• Cannot differentiate whether response to therapy is positive or
negative
![Page 8: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/8.jpg)
• Periodontal disease is localised and multifactorial.
Periodontal
pathogens
Host response
behaviouralsystemic
Genetic
![Page 9: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/9.jpg)
• Consideration should be given microbiologic
immunologic
systemic
genetic
behavioural factors
in addition to clinical and radiographic parameters.
![Page 10: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/10.jpg)
• The focus is now disease prevention, early discovery, and intervention to minimize treatment, thus enabling the most desirable outcomes.
• Diagnostic modalities available to clinicians today expand greatly on the foundation of a comprehensive visual assessment, which has been and will be the cornerstone of the diagnostic process.
![Page 11: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/11.jpg)
Classification
• Aids used in clinical diagnosis
i. Conventional probes – regular examination
ii. Millimeter probes – for gingival bleeding
iii. Pressure sensitive probes
Other clinical diagnostic aids
i. Filter papers- for measuring GCF
ii. Periotron 6000- for measuring GCF
iii. Olfactometer- for mouth odors
iv. Mobilometer- for tooth mobility
v. PSR- for faster screening and recording of PD
![Page 12: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/12.jpg)
• Aids used in radiographic diagnosis
i. IOPA radiographs
ii. Ortho-pantomograph
iii. Xero-radiography
• Aids used in microbial diagnosis
i. Direct examination
a. Light microscopy
b. Dark field microscopy
ii. Culture tests
a. Aerobic culture
b. Anaerobic culture
![Page 13: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/13.jpg)
• Aids used in immunological diagnosis
i. Immunofluorescense- direct and indirect
ii. Polymerase chain reaction
iii. Latex agglutination
iv. Flow cytometry
v. ELISA• Biochemical diagnosis
i. Studies for prostaglandins
ii. Studies for collagenase• Other diagnostic aids
i. study casts
ii. FSEIA
iii. N-benzoyl-DL- arginine 2- naphthylamide (BANA)
![Page 14: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/14.jpg)
Advanced diagnostic aids
• Advanced periodontal probes
i. Automated controlled force probes
ii. Thermal periodontal probes• Advanced diagnostic aids in periodontal radiography
i. Digital radiography
ii. Substraction radiography
iii. Digital substraction radiography
iv. Transmission radiography
v. Magnetic resonance imaging
vi. Computerized tomography
vii. Nuclear medicine bone scan
![Page 15: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/15.jpg)
• Advanced diagnostic aids in microbiologic analysis
i. Advances in culturing technique
ii. Advances in immunodiagnostic methods
iii. Advances in enzymatic methods
iv. Advances in nuclear biology- PCR and DNA probes
• Advanced diagnostic aids in charting the host response
i. Assessment of inflammatory mediators and products
ii. Assessment of tissue breakdown products
iii. Assessment of host derived enzymes
![Page 16: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/16.jpg)
• Advanced diagnostic aids to determine periodontal disease activity
i. Crevicular contents• Products of bacteria• Products of host cells and host immunity
ii. Markers in peripheral blood• Neutrophil functional profile• Monocyte responsiveness to LPS• Circulating antibodies to plaque bacteria
iii. Detection of specific periodontal pocket bacteria
a. DNA probes
b. BANA hydrolysis
c. Antibody techniques
![Page 17: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/17.jpg)
Advances in clinical diagnosis
• Degree of gingival inflammation
a. redness and swelling
b. gingival bleeding• Gingival bleeding ∞ plaque accumulation
gingival inflammation
(Greenstein G et al 1981)
size of inflammatory infiltrate
probability of losing attatchments
(Lang et al.1991)
![Page 18: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/18.jpg)
Gingival temperature
• Thermal probes are sensitive diagnostic devices for measuring early inflammatory changes in gingival tissue.
(Kung et al 1990)• Commercially available system periotemp probe• Individual temperature differences are compared with those
expected for each tooth and higher temperature pockets are signaled with a red emitting diode.
• Subgingival temperature at diseased sites is increased as compared to normal healthy sites
![Page 19: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/19.jpg)
• Elevated subgingival temperature ∞ attatchment loss
& elevated propertions of
periodontopathic bacteria
( Haffajee et al.)
![Page 20: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/20.jpg)
Periodontal probes
• Most widely used• Clinical assessment of connective tissue destruction in
periodontitis• Gold standard – recording changes in periodontal status• Probing depth is measured from the free gingival margin
(FGM) to the depth of the probable crevice.• not the most objective measure of loss of periodontal tissues
![Page 21: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/21.jpg)
• CAL is a more objective measure of loss of Existing periodontal support.
• CAL also does not give any indication of current disease activity.
• When interpreting the PD and CAL measurements made with conventional periodontal probes, it is important to consider that these values depend on the inflammatory state of the tissues.
![Page 22: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/22.jpg)
Classification of periodontal probes depending on generation.1.First generation probes:(conventional probes)Conventional manual probes that do not control probing force or pressure and that are not suited for automatic data collection.eg: Williams periodontal probe CPITN probe UNC-15 probe University of Michigan’O’ probe Goldman Fox probe Glickman probe Merritt A and B probe
![Page 23: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/23.jpg)
2.Second generation probe:
(Constant force probe)
Introduction of constant force or pressure sensitive probes allowed for improved standardization of probing.
e.g.: Pressure sensitive probe
Constant pressure probe
3.Third generation probe:(Automated probes)
Computer assisted direct data capture was an important step in reducing examiner bias and
also allowed for generation probe precision.
e.g.: Toronto probe
Florida probe
Interprobe, Foster Miller probe.
![Page 24: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/24.jpg)
4.Fourth generation probes:(Three dimensional probes)
Currently under development, these are aimed at recording sequential probe positions along a gingival sulcus.
An attempt to extend linear probing in a serial manner to take account of the continuous and three dimensional pocket that is being examined.
5.Fifth generation probe:(Noninvasive)
Three dimensional probe.
Basically these will add an ultrasound to a fourth generation probes.
If the fourth generation can be made, it will aim in
addition to identify the attachment level without
penetrating it.
e.g.: Ultra sonographic probe.
![Page 25: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/25.jpg)
Florida probe:
Tip is 0.4mmSleeve- edge provides reference
to make measurementsCoil Spring; provides constant probing forceComputer for data storage.
![Page 26: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/26.jpg)
![Page 27: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/27.jpg)
• Disadvantages of Florida probe.Lack of tactile sensitivityFixed probing forceUnderestimation of deep periodontal pockets.Other electronic probes:Improvised Florida PASHA probeInterprobePerioprobeFoster Miller probeToronto Automated ( difficult to reproduce patient head
position and in 2nd and 3rd Molar area.)
![Page 28: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/28.jpg)
PSR system
• To screen dental patients to facilitate the detection of mild forms of periodontal diseases and to identify individuals who have previously undetected periodontitis.
• It is designed for general dental practitioner to identify the patient’s requiring periodontal care and to determine the type of care required.
![Page 29: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/29.jpg)
• Based on the worst site per sextant.• If one or more sextants show significant signs of disease,
clinician is advised to do a complete periodontal examination and charting
![Page 30: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/30.jpg)
Advances in Radiographic Assessment
• Dental Radiography are traditional method to assess destruction of alveolar bone.
• Problems with conventional Radiography: Variation in projection geometry Variation in contrast and density Masking by other anatomic structures. They are very specific but lack sensitivity.
![Page 31: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/31.jpg)
Digital Radiography
• Computerized images.• Image property almost equal to conventional radiographs• 1/3rd to half reductions in radiation dose.
![Page 32: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/32.jpg)
Substraction Radiography
• Well established in medicine• Introduced in Periodontal diagnosis• Principle: Serial radiographs converted to digital images
superimposed composite image– Quantitative changes• Changes in density and volume of bone
a. can be detected as lighter areas (bone gain)
b. Dark areas (bone loss)
![Page 33: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/33.jpg)
• Perfect accuracy at a lesion depth of 0.49 mm
( Grondhal et al 1988)
• Limitations: needs paralleling technique and accurate
superimposition.
![Page 34: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/34.jpg)
Diagnostic Subtraction Radiography
• Positioning device during film exposure
specialized software designed for digital image subtraction
using conventional personal computers
• Applies an algorithm that corrects for the effects of angular alignment discrepencies and provides some degree of flexibility in imaging procedure
![Page 35: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/35.jpg)
Computer Assisted Densitometric Image Analysis(CADIA)
Video cameras
measures
light transmitted through Radiographs
signals from camera
converted into
gray scale images
camera interfaced with image processor
+ computer
![Page 36: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/36.jpg)
• Offers objective method for following alveolar bone density changes quantitatively over time.
• Higher sensitivity• High degree of reproducibility• accuracy
![Page 37: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/37.jpg)
COMPUTED TOMOGRAPHY
• Computed Tomography scanning is widely used in the evaluation of the implant patient
![Page 38: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/38.jpg)
• A thin fan beam of X-Rays rotates around the patient to generate in one resolution a thin axial slice of the area of interest.
• Multiple overlapping axial slices are obtained by several revolution of the X-ray beam until the whole area of interest is covered.
• With the help of a computer and sophisticated Algorithms these slices are then used to generate a three dimensional digital map of the jaw which help in evaluation of the implant patient.
![Page 39: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/39.jpg)
• Specialized software can be used to generate appropriate views that best depict the dimensions of the jaws and the location of important anatomic structures.
• DENTAL VIEWS OBTAINED FROM A CT SCAN INCLUDE:-
» 1. AXIAL» 2. PANORAMIC» 3. CROSS-SECTIONAL. Views of the Jaws.
![Page 40: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/40.jpg)
![Page 41: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/41.jpg)
ADVANTAGES of Computed Tomography
1. True cross sections offer a precise and detailed evaluation of the height and width of the ALVEOLAR RIDGE.
2. The images can be adjusted and printed without magnification, facilitating measurements directly on the prints or films with standard rulers.
3. Various anatomic structures can be visualized and analyzed at all the Coordinate axis.
1. Superioinferior 2. Anteroposterior 3. Buccolingual
![Page 42: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/42.jpg)
– Images of the entire arch several edentulous areas can be visualized with single examination.
– The Bone and soft tissue contrast and resolution are excellent for the diagnostic task.
![Page 43: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/43.jpg)
DISADVANTAGES of Computed Tomography specialized equipment and setting. Radiologists and Technicians need to be
knowledgeable of the anatomy, anatomic variants and pathology of the jaws as well as considerations pertinent implant treatment planning.
higher radiation dose to the patient as compared to other modalities used during implant treatment planning.
it delivers radiation to whole arch. Metallic Restorations can cause ring artifacts that
impair the diagnostic quality of the image, it is challenging to the patients having heavy metallic restored dentition.
![Page 44: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/44.jpg)
CONE-BEAM COMPUTED TOMOGRAPHY
• Cone-Beam Computed Tomography (CBCT) is a new imaging modality that offers significant advantages for the evaluation of implant patients.
![Page 45: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/45.jpg)
• The 3- D CBCT images canbe combined with high precision 3D visible light (photographic) surface images to create a virtual patient that accurately displays both hard and soft tissue structures.
• This multi- modal image image visualization enables a treatment platform that allows assessment of the patient’s present condition, planning and stimulation of treatment options, progress monitoring and evaluation of outcomes.
![Page 46: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/46.jpg)
Advances in Microbiologic Analysis
• Strong evidence for actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Tannerella forsythia (Tf).
• Other organisms that are thought to have etiologic role are Camphylobacter rectus, Eubaterium nodatum, Fusobacterium nucleatum, Peptostreptococcus micros, Prevetolla intermedia and Prevetolla nigrescens, Trepenoma Denticola
![Page 47: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/47.jpg)
• Microbiologic tests( that can identify)
a. Can support diagnosis of various Periodontal disease.
b. Can tell about initiation & progression
c. Active & Inactive
d. Can also be used to monitor Periodontal therapy
whether it is suppressed/ eradicated. • Several studies
(-)nce of Pathogens better periodontal health
(+)nce of pathogens periodontal disease
![Page 48: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/48.jpg)
• Bacterial culturing• Direct microscopy• Immunodiagnostic methods• Enzymatic methods• Diagnostic assays based on molecular biologic techniques
![Page 49: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/49.jpg)
Bacterial culturing
• Historically, widely used in characterizing composition of subgingival microflora.
• Plaque sample anaerobic culture
selective non-selective
Advantage: 1. clinician can obtain absolute or relative count
2. invitro method for antibiotic susceptibility
![Page 50: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/50.jpg)
• Limitations :
i. Culture can grow only live bacteria.
ii. Sensitivity is low
iii. Requires sophisticated equipment and experienced personnel.
iv. Time consuming
v. expensive
![Page 51: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/51.jpg)
Direct microscopy
• Dark field
Direct microscopy alternative to culture methods
• Ability to assess morphology
& motility in plaque.• Dark field microscopy seems an unlikely candidate as a
diagnostic test of destructive periodontal diseases.
![Page 52: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/52.jpg)
Immunodiagnostic methods
• It employs Antibody
that recognize
specific bacterial antigen• Various procedures:
a. Direct & indirect immunofluorescence assays
b. Flow cytometry
c. ELISA
d. Membrane assay
e. Latex agglutination
![Page 53: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/53.jpg)
Immunofluorescence assay
• Direct IFA: AB conjugated with Fluorescein marker + Bacteria ( Antigen) = Immuno complex
• Indirect IFA: Primary AB + Bacteria= Immune Complex+ Secondary Fl conjugated AB
![Page 54: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/54.jpg)
Direct IFAIndirect IFA
![Page 55: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/55.jpg)
Cytofluorography/ Flow cytometry
Bacterial cells+ species specific AB
+ Secondary FL Conjugated AB Introduced in flowcytometer
Bacterial cells is separated into
single cell suspension-
passes through the tube
Cells identified by lasers
![Page 56: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/56.jpg)
• merits :
rapid identification
labels bacterial cells species specific antibody
fluorescin conjugated secondary
antibody• Demerits:
sophistication
expensive
![Page 57: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/57.jpg)
ELISA= Enzyme Linked Immunosorbent Assay
Similar AB and Antigen reaction, but the fluorescence is read using a photometer.
Evalusite: commercially available kit to detect Aa,Pg and Pi.
![Page 58: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/58.jpg)
![Page 59: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/59.jpg)
Well with precoated antibody + Sample to be tested= immune complex
Specific antigen bind to the antibody + Secondary antibody added.
Immunofloresence dye bound to secondary antibody
Substrate added which changes the color of the solution
Amount of florescence checked by photometer (450nm)
![Page 60: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/60.jpg)
• Latex Agglutination Test
Latex beads coated with species specific AB when beads come in contact with specific species in sample they bind and agglutination occurs clumping of beads is visible test positive.
Advantages:Simple and Rapid testing
Higher sensitivity and specificity.
![Page 61: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/61.jpg)
Enzymatic methods
• Bacteria release specific enzymes. Certain group of species share common enzymatic profile.
e.g. Tf, PG, Td, and Capnocytophaga species release trypsin like enzyme.
• Enyme hydrolyses specific substrate (BANA) release cromophore ( B-naphtalamide) Cromophore changes color on addition of a substance( fast garnet)
![Page 62: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/62.jpg)
Perioscan is a popular diagnostic kit uses BANA reaction.
Disadvantage:
May be positive in clinically healthy site
Cannot detect disease activity
Limited organisms detected
Other pathogens may be present if it’s negative.
![Page 63: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/63.jpg)
Molecular Biology Techniques
• Basic Principle: Analysis of DNA, RNA and protein structure.
Hybridization: Pairing of complimentary strands of DNA to produce a double stranded DNA
.
Nucleic acid probe: is a known DNA/RNA which is synthesized artificially and labeled with a enzyme or a radioisotope for detection when placed in a plaque sample
![Page 64: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/64.jpg)
DNA Probe: uses a segment of a single stranded DNA, labeled with a enzyme of a radio isotope, that is able to hybridize to a complimentary nuclei strand, and thus detect presence of target microorganism.
![Page 65: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/65.jpg)
DNA Probe
Whole genomic: Targets the whole DNA strand rather then a specific sequence or gene.
High chances to cross react with non target microorganism
Lower sensitivity and specificity.
![Page 66: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/66.jpg)
Oligonucleotide probes: target variable region of 16sRNA or a specific sequence in the DNA strand.
Higher sensitivity and
specificity.
![Page 67: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/67.jpg)
• Checkerboard DNA-DNA Hybridization Technology:
Developed by Socransky et.al.
40 bacterial species can be detected using whole genomic digoxigenin-labeled DNA probes.
Large number of samples can be tested and upto 40 oral species detected with a single test.
![Page 68: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/68.jpg)
Advantages of DNA probes as compared to bacterial culturing.
1. More sensitive and specific2. Requires as less as 104 cells of each species to be
detected.3. Multiple species detected with a single test4. Does not require viable bacteria5. Large number of samples can be assessed.Disadvantage:6. Expensive 7. Expert personnel to carry out the test8. Not easily available
![Page 69: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/69.jpg)
• Polymerase Chain Reaction (PCR):Involves amplification of a region of DNA by a primer specific to
the target species.
If there is amplification then it indicates the presence of the target species in the sample.
![Page 70: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/70.jpg)
Advantages:1. High detection limit. As less as 5- 10 cells can be
amplified and detected.2. Less cross reactivity under optimal conditions3. Many species can be detected simultaneouslyDisadvantage:4. Small quantity needed for reaction may not contain the
necessary target DNA5. Plaque may contain enzymes which may inhibit these
reactions.
![Page 71: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/71.jpg)
Advances in characterizing the host response
• Diagnostic tests have been developed that add measures of the inflammatory process to conventional clinical measures.
• These diagnostic tests gives information about
pattern of destruction
current activity of disease
rate of disease progression
extent & severity of further breakdown
response to therapy
![Page 72: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/72.jpg)
• So, knowing about disease destructive process-
clinician can individualise their therapeutic approach,
and customize the treatment.• Source of samples may be; GCF, Saliva, or Blood.• GCF is most commonly used, where as saliva is recently been
researched.• Assessment of Host response
i. Inflammatory mediators and products
ii. Host derived enzymes
iii. Tissue breakdown products
![Page 73: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/73.jpg)
Inflammatory mediators & products:
• Cytokines- potent local mediators of inflammation
produced by variety of cells• GCF- TNF α
IL-1
IL-6
IL-8• Good correlation with disease status and severity but not
disease progression
![Page 74: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/74.jpg)
• In untreated periodontitis- concentration of PGE2 increased during active phase of periodontal destruction.
• Cytokines ∞ disease severity & status• PGE2 ∞ active phase of periodontal disease
![Page 75: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/75.jpg)
Host derived enzymes:
• Matrix components are dissolved by either
a. ECM metalloproteinase dependent
b. Plasmin dependent cleavage reactions
• Proteases and enzymes involved in these processes may be used as diagnostic aids.
![Page 76: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/76.jpg)
• Aspartate aminotransferases• Collagenase• β-glucuronidase• Lactate dehydrogenase• Arylsulfatase• Elastase• Alkaline phosphatase
![Page 77: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/77.jpg)
Tissue Breakdown Products
• Hydroxyproline• Glycosaminoglycans
![Page 78: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/78.jpg)
For periodontal diagnosis, the ideal diagnostic test should be • Quantitative.• Highly sensitive method capable of analyzing a single
periodontal site in health as well as disease.• Reproducible.• Highly specific.• Simple to perform.• A rapid, one or two stage procedure.• Non-invasive.• Versatile in terms of sample handling, storage and transport.• Amendable to chairside use.• Economical. ( Chapple 1997)
![Page 79: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/79.jpg)
Chairside periodontal test kits can be categorized as• Microbiologic tests• Biochemical test kits• Genetic kits
Microbiological test kits
The bacteriological tests
(Microscopy, Culture, Omnigene, Affirm DP and Evalusite) are mainly aimed at spirochetes, Aa, Pg and Pi.
Microbial tests can also be used to monitor periodontal therapy directed towards the suppression or eradication of periodontopathogenic microorganisms
![Page 80: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/80.jpg)
Omnigene
• DNA probe systems
• available for the detection of A. actinomycetemcomitans, P. gingivalis, P. intermedia, F. nucleatum, C. rectus, T. denticola
and E. corrodens.
![Page 81: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/81.jpg)
Evalusite
• Evalusite is a kit that employs a novel membrane-based enzyme immunoassay for the detection of three putative periodontopathogens: Aa, Pg and Pi.
• a pink spot is displayed if the test organism is present.• The main weaknesses of this test kit reside in• 1) the assumption that the three detected organisms are
causing disease;• (2) it is a multistage test; • (3) it has a subjective calorimetric end point and (4) there is no
permanent record of the results.
![Page 82: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/82.jpg)
PerioScan®
• Perioscan is a diagnostic test kit that utilizes the BANA (N-benzoyl-DL-arginine-2-naphthylamide)-hydrolysis reaction,
• developed to detect bacterial trypsin-like proteases in the dental plaque
![Page 83: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/83.jpg)
Biochemical test kits
• Perio 2000- to evaluate VSCs• Prognos- Stik- to detect elevated levels of MMPs• Perio- Check- to measure neutral protease activity• PerioGard- to detect AST• Pocket Watch- simple method for AST
![Page 84: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/84.jpg)
Genetic test kits
• Various gene polymorphisms are considered to be risk factors for the initiation or progression of periodontal disease.
• In 1997, Kornman et al. found an association between the polymorphism in the genes encoding for interleukin-1α and interleukin-1β and increased severity of periodontitis.
• PST® genetic susceptibility test - the first and only genetic test that analyzes two interleukins (IL-1α and IL-1β) genes for variations. IL-1 genetic susceptibility may not initiate or cause the disease but rather may lead to earlier or more severe disease
![Page 85: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/85.jpg)
Salivary diagnostics
• OralDNA® Labs has developed a salivary test, the
MyPerioID® PST®, that identifies a patient’s genetic susceptibility and inherent risk to periodontal disease by evaluating their interleukin-1 (IL-1) gene cluster,
• MyPerioPath®, that identifies the type and concentration of 13 pathogenic bacteria known to cause periodontal disease.
![Page 86: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/86.jpg)
Conclusion
• In periodontology, the success of any treatment is dependent upon the accuracy of the initial diagnosis.
• At present, the majority of chronic periodontitis cases can be adequately managed using existing diagnostic methodology, although it is clearly more desirable to be able to diagnose “active disease” as it occurs, rather than months later.
• However, the clinician must ensure that the use of such tests will benefit the patients in terms of both the value of diagnostic data obtained and the cost in time and money.
![Page 87: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/87.jpg)
References
• Carranza’s Clinical Periodontology 10th and 11th edition.• Armitage GC, Jeffcoat MK, Chadwick DE: longitudinal
evaluation of elastase as a marker for the progression of Periodontitis, J Periodontol 1994; 65:120.
• Beck JD: Issues in assessment of diagnostic tests and risk for Periodontal diseases, Periodontol 2000;7:100
![Page 88: Advanced diagnostic aids khushbu](https://reader036.vdocuments.us/reader036/viewer/2022062418/554b2ef0b4c905da088b4e59/html5/thumbnails/88.jpg)
Thank you