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Perio Exam 2 > Efficacy of Diagnostic Techniques > Flashcards

Efficacy of Diagnostic Techniques Flashcards

1
Q

Microbiologic Testing

A 
Expensive and well-training personnel required 
 Bacterial culturing
 Direct microscopy
 Immunodiagnostic methods
 Enzymatic methods
 Molecular biology techniques
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2
Q

Assessment of the Host Response

A

Biochemical analysis as part of periodontal diagnosis
Source of samples:
GCF, saliva, and serum (blood)

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3
Q

Genetic Analysis (only for research)

A

There is a genetic susceptibility to periodontitis

Gene polymorphism as a risk marker for periodontitis

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4
Q

Probe Penetration

A

Lack of sensitivity and reproducibility
Probing depth: gingival inflammation, insertion force, placement and angulation, size, probing technique, probe calibration, presence of subgingival calculus, overhanging restorations

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5
Q

CAL

A

Poor reliability and reproducibility

Limited practical value

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6
Q

Radiographic Examination

A

Limited sensitivity in small bone change
– Changes in bone can be identified by eye only after 30% to 50% of the bone mineral has been lost (Subtraction
radiography: detect bone density change as low as 5 %)
No value in evaluating disease activity or progression

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7
Q

Radiographic exam: simply a measurement of the

A

past damage of a disease process. +- 1mm for probing depth – this will vary based on person to person.

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8
Q

Ultrasonic periodontal probe uses a

A

hollow tapered tip that is filled with water for coupling of the ultrasonic beam into the tissues (non-invasive)

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9
Q

Cone-beam Computed Tomography: Conventional radiographs (PA; Pano) are very

A

specific, but lack sensitivity

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10
Q

Cone-beam Computed Tomography: Recently, dental CBCT has been introduced in periodontology for the detection of

A

periodontal defects in in vitro settings

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11
Q

Cone-beam Computed Tomography: CBCT is promising for periodontal applications, especially for

A

intrabony defects, dehiscence and fenestration defects, periodontal cysts, furcation defects and thickness of palatal masticatory mucosa

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12
Q

The sensitivity of a diagnostic test refers to the

A

probability of the test being positive when the disease is truly present

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13
Q

The specificity of a diagnostic test refers to the

A

probability of the test being negative when the disease is not present

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14
Q

Fastidious microorganism – microorganisms that will grow only if

A

special nutrients are presence

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15
Q

Assess for antibiotic susceptibility of microbes while

A

culturing

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16
Q

Some putative pathogens are fastidious and

A

difficult to culture

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17
Q

Sensitivity is low: detection limits for selective and nonselective media average

A

104 to 105 bacteria

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18
Q
  1. Direct Microscopy
A

Alternative to culture methods
Dark-field or phase-contrast microscopy
Morphology and motility of bacteria in a plaque sample

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19
Q

Problem with direct microscopy:

A

Most of the main putative perio pathogens are non-motile (so it is difficult to identify)

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20
Q
  1. Immunodiagnostic Methods
A

Use Ab that targets specific
bacterial Ag
Direct and indirect - immunofluorescence microscopic assay

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21
Q 
immunofluorescent 
microscopic assay (IFA)
A 
Able to identify pathogens 
   using a plaque smear
Used mainly to detect Aa and Pg
Comparable to bacterial culture
Does not require viable bacterial cells
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22
Q
  1. Immunodiagnostic Methods: Cytofluorography (flow cytometry)
A

complexity and cost prevent its wide use

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23
Q

Immunodiagnostic Methods: Enzyme-linked immunosorbent assay (ELISA)

A 
Used primarily to detect serum antibodies to periodontal pathogens
Membrane immunoassay (EvalusiteTM): chairside use to detect Aa, Pg, and Pi (detection limit of 105 for Aa and 106 for Pg)
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24
Q

Immunodiagnostic Methods: Latex agglutination

A

Based on the binding of protein to latex: latex beads are coated with species-specific antibody
Currently these assays only for research purposes

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25
Q
  1. Enzymatic Methods
A

Several putative periodontal pathogens such as Pg, Tf, and Aa possess in common a trypsin-like enzyme that hydrolyzes a substrate N-benzoyl-DL-arginine-2-naphthylamide (BANA).
Chair-side kit (PerioscanTM) was available in the 1990s
Inability to distinguish between individual bacteria
- It may be positive at clinically healthy sites
Negative result does not rule out the presence of other important periodontal pathogens

26
Q

16s rRNA is like a finger print:

A

comparing certain locations on a 16s rRNA molecule with a database of known organisms allows the identification of organisms whose 16s rRNA signature is known.

27
Q

Real-time PCR is also known as

A

The quantitative real time polymerase chain reaction (qPCR). The Real-Time-PCR enables both detection and quantification.

28
Q

Nucleic acid probes

A

Synthesized and labeled DNA (20-30 nucleotides)
Genomic DNA probe (whole DNA strand): significantly ↓ in sensitivity and specificity due to cross-reactivity to non-target microorganisms
16S rRNA – oligonucleotide probes (high sensitivity and specificity

29
Q

Checkerboard DNA-DNA hybridization

A

Whole genomic digoxigenin-labeled DNA
Up to 40 oral bacterial species in a single test
Not been generalized for diagnostic purpose

30
Q

PCR

A

High sensitivity and specificity for the identification of target pathogens
PCR lower detection limit: 25-100 cells (Culture: 104-105 cells)
Unable to quantify pathogens accurately in clinical samples

31
Q

Real-time PCR

A

Real-time PCR: good correlation between the fluorescent signal measured and the number of bacterial cells been used
Expensive and sophisticated technology in real-time PCR

32
Q

Hypophosphatasia (OMIM 146300, 241500, 241510) is an inherited disorder characterized by

A

defective bone and teeth mineralization and deficiency of serum and bone alkaline phosphatase (AP) activity.

33
Q

urinary phosphoethanolamine (PEA) is increased in

A

hypophosphatasia.

34
Q

urinary inorganic pyrophosphate (PPi) level is

A

also increased in hypophosphatasia

35
Q

Collecting GCF

A

Paper strips placed within the crevice for 30 seconds
Fluid volume can be quantified by Periotron®
Captured samples may not represent the entire periodontium
Selection of the teeth and sites is often difficult

36
Q

GCF

Currently > 65

A

components of GCF have been evaluated
Host-derived enzymes and their inhibitors
Byproducts of tissue breakdown
Inflammatory mediators and host-response modifiers

37
Q

Intracellular destruction enzymes

A

Possible markers of active periodontal destruction

Released from dead or dying PMN/Neutrophils from periodontium

38
Q

Aspartate amino-transferase: released during

A

tissue destruction (cell death)

39
Q
  • Alkaline phosphatase: a membrane-bound
A

glycoprotein involved in maintenance of alveolar bone

40
Q
  • β-glucuronidase: a
A

lysosomal enzyme degrades proteoglycans and ground substance

41
Q
  • Elastase: a
A

proteolytic enzyme found in lysosomal granules of neutrophil

42
Q

Extracellular destruction enzymes

A

Associated with the activity of matrix metalloproteinases

Produced by inflammatory, epithelial and connective tissue cells

43
Q

Aspartate aminotransferase (AST)

A

Periogard Periodontal Tissue MonitorsTM (chair-side test kit)
A marked elevation in AST levels in GCF from sites with severe gingival inflammation
Inability to discriminate between sites with severe inflammation with or without attachment loss

44
Q

Alkaline phosphatase (ALP)

A

ALP in GCF are higher in diseased then healthy sites alkaline phosphate in disease.

45
Q

β-glucuronidase (βG)

A

Elevated βG in GCF from sites with severe periodontal disease
High sensitivity and specificity when related to occurrence of clinical attachment loss
Good predictor for future periodontal breakdown

46
Q

Elastase

A

Periocheck® (chair-side test kit)

Positive correlation of elastase in GCF with clinical attachment loss

47
Q 
Matrix metalloproteinases (MMPs)
Secreted by
A

fibroblasts and macrophages

48
Q

MMPSL: Responsible for

A

remodeling and degradation of ECM components

49
Q

MMPS: Regulated by

A

tissue inhibitors of MMPs (TIMPs)

periodontal ECM

50
Q

MMPS: High MMP levels in GCF are at significantly greater risk for

A

progression of periodontitis

51
Q

GCF MMPs level reduces in response to

A

treatment

52
Q

Destruction of collagen

The ECM of the periodontium is composed of

A

collagen (predominant), proteoglycan (versican, decorin, biglycan, syndecan) and non-collagen proteins (elastin, fibronectin, laminin, osteocalcin, osteopontin, bone sialoprotein, osteonectin, and tenascin)

53
Q

Elevated levels of hydroxyproline (breakdown from collagen), glycosaminoglycans (from matrix degradation) and osteocalcin & type I collagen (from alveolar bone destruction) can be found in the

A

GCF from sites with periodontitis

54
Q

Traditional immunoassays analyze only a

A

single cytokine at a time

55
Q

Cytokines

A

inflammatory mediator

56
Q

Bio-Plex Cytokine Assay

A

Incorporates novel technology using color-coded beads, permits the simultaneous detection of up to 100 cytokines in a single well of a 96-well microplate
Are multiplex bead-based assays designed to quantitate multiple cytokines in tissue fluid including GCF

57
Q

OPTICAL SPECTROSCOPY

Infrared (IR) Spectroscopy

A

Vibrating covalent bonds of organic molecules absorb a characteristic wavelength of IR light
The wavelength of light absorbed depends on the nature of the covalent bond (e.g., C=O and N–H), the type of vibration (e.g., bending and stretching), and the environment of the bond
The spectrum of absorbed light can be used to establish a molecular fingerprint of a tissue or fluid

58
Q

Diagnosis of Periodontitis Based on IR Spectra of GCF

A

Measure the total contents of GCF
IR spectroscopy can be used to characterized GCF from healthy, gingivitis, and periodontitis sites
Vibrations of peptide groups: C=O stretching (amide I band) and N–H bending (amide II band)
IR spectroscopy of GCF is reagent free, requiring only small sample volumes, requiring minimal training for operator

59
Q

Near Infrared (NIR) Spectroscopy

A

Measure of oxygen saturation of the tissues
The wavelength region 500 to 600 nm is dominated by the absorption from oxygenated hemoglobin (HbO2) and deoxygenated hemoglobin (Hb)
Tissue oxygenation at periodontitis sites significantly ↓ as compared to gingivitis and healthy control sites

60
Q

Why Saliva for periodontal disease testing?

A

Abundant fluid and easy to collect and store
Highly enriched content of disease biomarkers
- Such as Sjogren’s syndrome and oral cancer

61
Q

Salivary Occult Blood Test (SOBT) –

A

available in Japan
A poor periodontal status: Subjects having ≥15% of teeth with BOP or ≥ 1 tooth with PD ≥ 4mm
A paper strip containing gold-labeled anti-human hemoglobin monoclonal antibody was dipped into saliva sample (Positive: ≥ 2 μg/ml human hemoglobin)
Sensitivity:0.72; Specificity: 0.52

62
Q

The SOBT may offer a simple screening method for periodontal status when

A

a thorough periodontal examination is not possible, although it is not sufficiently specific to be a reasonable substitute for a periodontal examination

Perio Exam 2 (20 decks)

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