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LABORATORY
INVESTIGATIONS
Contents
•   Introduction
•   Tests done by dentists
•   Hematological
•   Urine Analysis
•   Biochemistry :Renal function tests
                Tests – diseases of bone
                Liver function tests
               Lipid Analysis
•   Immunological investigations
•   Histopathology, cytology and immunofluorescent studies Microbiology
•   Radiological
•   Conclusion
•   References
INTRODUCTION
• Diagnosis & identification - disease by careful investigation o
  patients signs, symptoms and history

• Carefully crafted history and physical examination -
  satisfactory – diagnosis

• Times when more information is required through the use of
  diagnostic tests.

• Clinical and/or lab data must be used to distinguish between
  different diagnoses.
importance of laboratory
            investigations

• laboratory tests - important in assisting & management
  of the patient during treatment of disease besides
  diagnosis.

• Used-
1. screen - disease in asymptomatic individual
2. to establish or exclude presence of diseases in
   symptomatic patients
3. assist the practitioner in the management of the patient.
important characteristics of a
                 laboratory test

•   Accuracy
•   Cost
•   Interfering factors
•   Precision
•   Reference range
•   Sensitivity
•   Specificity
TESTS ADVISED BY DENTISTS
      Acc. Sonis, Fazio & Fang
Depending upon the specimens
            submitted
1. Hematological

2. Biochemical

3. Immunological

4. Histopathological – histological & Immunofluorescent

5. Microbial

6. Radiological
Depending upon the organs involved

•   Heart                           •   Skin
•   Vascular system                 •   Eye
•   Respiratory system              •   Bones & joints
•   GIT                             •   Skeletal muscle
•   Liver, gallbladder & pancreas
                                    •   CVS
•   Kidney & genitourinary
                                    •   Mental illness
•   Female genital
                                    •   Endocrine system
•   Breast
                                    •   Infectious diseases
                                    •   Immunological diseases
Laboratory investigation
 Biochemistry :
 Urine analysis
 Salivary chemistry
 Histopathology, cytology,
  Immunofluorescent studies
 Serology & immunohistochemistry
 Radiology
BLOOD CHEMISTRY TESTS

  A. . Estimation of electrolytes

  B. . Renal function

  C. .Liver function

  D. .Thyroid Function

  E. . Metabolic bone disease

  F. . Other tests .
A. Estimation of electrolytes

• Maintain fluid levels inside & outside cells

• Osmotic gradient – nerve condtn. Muscle func,hydration
   & maintaining Ph levels
1. SODIUM :

• Loss of sodium results      extracellular fluid volume

• Circulation, renal function, and central nervous system
  function.
• Normal : 136-145 m Eq/l
Feel weak,
confusion,
 paralysis,
 sezuires




                 Decrease
               consciousness,
              vomiting, fatigue,
              muscle weakness
2. Chloride
• Normal : 95 to 108 mEq/l.

• Important - maintenance of acid-base balance
3. Potassium
• determines neuromuscular irritability.

•    or     conc. impair - ability of muscle tissue contract

• Normal - 3.5 to 5.2 mEq/l.
4. Bicarbonate
• Bicarbonate–carbonic acid buffer - maintaining normal
  pH of body fluids.
• basis for assessing acid-base balance.
• Normal : 24 to 30 mEq/l.
Dentists

• Electrolytes -
• Maintain - Ph
• Proteins are denatured and digested
• Enzymes lose their ability to function and death may
  occur.

• possibly life threatening.
• could require hospitalization and intravenous
  electrolytes.
b. renal function test
1. blood urea nitrogen

•   Normal value of BUN in adults: 8 – 18 mg %.
                                   10 – 20 mg/dl
     sonis
                     symptom - >50 mg/dl
Clinical significance :
• Pre renal causes – reduced flow to kidney, shock, blood
  loss, dehydration, increased protein catabolism, injuries,
  burns, fever

• Renal causes – acute renal failure, glomerulonephritis,
  malignant hypertension, chronic renal failure, DM

• Post renal causes – urinary obstruction by stones,
  tumors
•   Decreased blood urea nitrogen –
•   Poor nutrition
•   High fluid intake
•   Excessive administration of i.v fluids
•   Liver damage
•   Late pregnancy, infancy, acromegaly.
2. creatinine

• Creatinine - catabolic product of creatinine phosphate - used for
  skeletal muscle contraction.

• excreted entirely - kidneys
• amount in blood - directly proportional to renal excretory
  function
• diagnose impaired renal function

• Normal range – 0.5 – 1.6mg% or 1.5 mg/dl
• symptom > 4mg/dl
INCREASED CREATININE LEVELS:
• Kidney disease



• Decreased: Pregnancy – NS

• decreased – improve function
Dentists
• O/E : Pale, fluid retention – edema, Inc IJP, asitics' -
  advice Investigations

• Anemic, Acidosis – K+ retention
• BT prolonged

• Rx – avoid anti platelet drugs –
       Aspirin NSAIDs, nephrotoxic drugs
• Consent – physician
C. LIVER FUNCTION
Serum bilirubin

Urine urobilinogen

Serum albumin – globulin

Serum alkaline phosphatase

AST & ALT
1.serum bilirubin



•   Bilirubin : excreted - intestines.
•   Intestines converted - colorless compound - urobilinogen.
•   Level of bilirubin elevates - excessive hemolysis
•   Inability of liver to eliminate bilirubin (liver damage or
    obstruction of bile duct)

• yellow color - product
• skin, mucous membranes, sclera of the eye, plasma and urine
• Excess bilirubin - associated - hepatitis
• Jaundice - total serum bilirubin rises

• Normal Value →         0.1 to 1.2 mg / 100 ml
                         5.1–17.0 micro mol/L
High values:

•   Hemolytic Anemia
•   Biliary obstruction
•   Hepatitis
•   Malignant Hepatic Disease
2. SERUM GLUTAMIC OXALOACETIC TRANSAMINASE (SGOT)

            SERUM GLUTAMIC PYRUVIC TRANSAMINASE (SGPT)

• enzymes -large amounts - liver, heart, kidney and skeletal muscle
• metabolism of amino acids and carbohydrates.

• NORMAL VALUES :

• SGOT also called Aspartate Aminotransferase 6 -25 I U/ L
• SGPT also called Alanine Aminotransferase 3– 26 I U/ L
• SGOT -sensitive indicator - myocardial necrosis, significant rises
  occurs

• Hepatitis & other liver disease + associated liver necrosis- SGOT
  and SGPT
screening for hepatitis b &c
• Detection of hepatitis B surface antigen:

• Hepatitis B virus (HBV) is a double stranded DNA spherical particle with
   a double shell. - referred as the Dane particle.


• Antigens which have so far been associated with HBV are as follows ;

• Hepatitis B surface antigen (HBsAg) : This was first noticed in the blood
   of an Australian aborigine hence earlier was termed as Australia antigen.
• Hepatitis B core antigen (HBcAg)

• Hepatitis B e antigen (HBeAg).
• Infection - HBV i- entry of virus through the skin or mucous
  membranes into the blood or body fluids.
Routes of transmission
• transfusion of blood or blood products.
• HBV contaminated needles, syringes or prickers.
• Close personal contact with a person with hepatitis B
• Contact with blood specimen (Laboratory personnel those who
  handle the specimen)
• Transmission by blood sucking insects or bed bugs.
Clinical Significance :
• detection of hepatitis – b surface
  antigen (hbsag) by direct enZyme
  linKed immunosorbent assay (elisa) :


• HBsAg formerly called Australia antigen is
• Reduce incidence of post-transfusion hepatitis,
  testing for HbsAg is mandatory for blood
  products intended for human use.
• Care – Rx – viral particles – saliva & other body
  secretions.
• Hepatitis A – SGOT, SGPT –
• Hepatitis B – SGOT,SGPT & LDH –
• Hepatitis C – SGOPT & SGPT

•   Dentists – possible signs – icterus, fever ../ H/O
•   Lab inv -
•   Viral hepatitis – infectious B
•   Exposed inf blood & saliva- Precautions
• Signs
• H/O: Alcohol abuse, hepatitis ,abnormal bleeding
• Features – improper function Liver – Def production
  clotting factors – PT elevated
• So investigation & physicians opinion – needed –
  precautions
3. serum albumin and globulin


• total protein
• important - coagulation, transport hormones,
  act on antibodies
• act - buffers with enzymes

• help - maintain osmotic pressure.
• Serum albumin - synthesized in liver
• globulins - produced - plasma cell
• Normal Values:

•   Total Protein   6.0 to 7.8 g / 100 ml or 6.0 to 8.3 gm/dL
•   Albumin             3.2 to 5.6 g / 100 ml
•   Globulin            2.3 to 3.5 g / 100 ml
•   A / G Ratio         1.5 to 1 to 2.5:1
Total proteins




High Albumin - rare but dehydration and shock.
Low Albumin : same as proteins.
High Globulin : Multiple myeloma, Nephrosis, Chronic infections,
  Collagen diseases, Liver diseases
Low Globulin : Burns and severe malnutrition.
5.Alkaline phosphatase
• high concentration - growing bone, bile, placenta.
• Normal level: 30 to 115 U/l.

Increased
A. In children
B. Osteoblastic bone disease
C. Hepatic disease / bileobstruction - stone, stricture,
   neoplasm.
D. Pregnancy.

Decreased:
• Hypophosphatasia, hypothyroidism, malnutrition.
Normal values and changes in two types of Jaundice

                                                      Hepatocellular            Uncomplicated
Tests                  Normal Values
                                                      Jaundice                  Obstructive Jaundice

Bilirubin
- Direct                      1. – 0.3 mg/dL          Increased                 Increased
-Indirect                     2. – 0.7 mg/dL          Increased                 Increased


Urine bilirubin        None                           Increased                 Increased

Serum albumin/Total    Albumin, 3.5-5.5 g/dL          Albumin decreased         Unchanged
protein                                               Total protein, 6.5-8.4
                                                      g/dL


Alkaline phosphatase   30-115 units /L                Increased (+)             Increased (++++)

Prothrombin time       INR of 1.0 – 1.4 After         Prolonged if damage       Prolonged if obstruction
                       Vitamin K, 10% increase        severe and does not       marked, but responds to
                       in 24 hours                    respond to parenteral     parenteral vitamin K
                                                      Vit K
        ALT, AST       ALT, 5-35 units/L, AST, 5-40         Increased in           Minimally increased
                                 units/L               hepatocellular damage,
                                                           viral hepatitis
D. THYROID FUNCTION
Reference values of thyroid
function test

Test              Range


TSH            0.5 - 4.7mU/L

T3             0.92-2.78nmol/L

T4             58-140 nmol/L

FT4            10.3-35pmol/L
HYPO THYROIDISM                             HYPER THYROIDISM

•    Catecholamines in LA +                  •   CNS depression – present
     stress in dental operatory              •   Administration of narcotic
•    Ppt thyroid Astorm                          analgesics increase risk
•    Characterised High fever,                   CNS depression& collapse
     psycosis, CNS depresssion,
     vomiting diaorrhea – risk
     CHF



    Pt asked for Heat & cold intolerance, wt gain /loss, change in appetite bowel
                    habits , muscle weakness and palpitations
E. Metabolic Bone Disease
•   Jaw lesions – radiographic examination
•   Systemic jaw diseases-

•   Pagets disease, FD, Primary & Sec.
    Hyperparathyroidism, Osteoporosis, MM, Osteogenic
    sarcoma or metastatic malignancy

•   Serum Ca, P,& alkaline phosphatase -
Serum Ca, P

• Serum cal I / serum phosphorus
• Measured : mg/dl
• serum Ca conc. X serum P conc. = constant

• 30 to 40 mg/dl = Normal adults
• 50 to 60 mg/ dl = growing children
•   GA & surgical procedures – cardiac arrythmmias, heart block

•   Hypercalcaemia - excessive skeletal calcium release, increased intestinal
    calcium absorption, or decreased renal calcium excretion.



•     serum Ca – hypo proteinmia, due decreased binding by serum protein &
    renal disease.
2. Alkaline phosphatase
• Oestoblasts
• enzyme - produced - small amounts –liver
• larger amounts by osteoblasts.
• active in bone formation and therefore is found at higher levels there.

•      result increased osteoblastic activity
• In association – obstructive liver disease, amyloid disease, leukemia
  & sarcoidosis
•   Normal values for serum Alkaline phosphatase
•   King Armstrong Units      – 4 to 13
•   Brodansky Units          - 1.5 to 4.5
•   International Units      - 30 to 85 IU
High Alkaline Phosphatase values :
•   Obstructive liver disease
•   Metastatic carcinoma involving bones
•   Hyper parathyroidism
•   Pagets Disease of Bone
•   Osteo malacia
•   Rickets
•   Amyloid Disease
•   Leukemia
•   Sarcoidosis

Low Alkaline Phosphatase :

• Hypophosphatasia
• Hypothyroidism
• Scurvy
Values                Serum calcium        Serum               Serum phosphates
                      mg/dl                phosphorus          Units /dl
                                           P/dl
Normal                8.8 to 10.5          2 to 5              1-4

Rickets               Normal               Decreased –         Increased 20 to 40 x
                                           Exc tetany          normal

Osteomalacia          Decreased            Decreased           Little if any change

Paget’s Disease       Normal               Normal              Occasionally
                                                               elevated

Hyperparathyroidism   Increased            Decreased           Increased 2 to 50 x
                                                               normal

Osteogenic            normal               Normal              Slightly increase
Imperfecta

Solitary Bone cysts   normal               normal              normal


Metatstatic Oseous    May be elevated      Normal              Normal / slightly
Disorders                                                      elevated
Tetany                7 mg ca/dl or less   Normal / elevated   Normal
G. Other Tests
1. Acid phosphatase
• Phosphatases active at pH 4.9 are present - high
   conc.

• prostate gland, erythrocytes, platelets,
  reticuloendothelial cells, liver, spleen, and kidney.

• Normal levels: 0.8 IU/l.

• Increased:
 carcinoma of the prostate
2. Serum amylase
• Normal level: 5 to 75 IU/l.
• Increased:
• Acute pancreatitis, pseudocyst of the pancreas,
• obstruction of pancreatic ducts (carcinoma, stone,
  stricture,duct sphincter spasm after morphine), and parotitis.
• Decreased:
• Acute and chronic hepatitis
• Pancreatic insufficiency,
• toxemia of pregnancy.
3. Serum lipase
Normal level: 0.2 to 1.5 units.



Increased:
• acute or exacerbated pancreatitis
• obstruction of pancreatic ducts- stone or neoplasm.
4. Serum cholesterol
• Normal Range : 160 to 300 mg/dl

• > 300 mg/dl : dietary reduction – animal fats, eggs &
  foods     cholesterol levels

• Risk – artherosclerotic CVD

• Elevated :
• hypothyroidism, obese; elderly diabetic & nephrotic
  syndrome
Check on
                         triglycerides




• HDL : good" cholesterol - removes excess cholesterol
  from the blood and takes it to the liver

• LDL: BAD: High levels- linked - increased risk of heart
  and blood vessel disease, inlcuding coronary artery
  disease, heart attack and death.
5. Creatine phosphokinase
              (CPK)
• Male: 50 to 180 IU/l, female: 50 to 60 IU/l.

• Myocardial infarction, trauma to muscle, malignant
• Hyperthermia muscular dystrophies, polymyositis,
  severe Muscular exertion (jogging), hypothyroidism
6. Lactate dehydrogenase
              (LDH)
• Normal level: 45 to 100 U/l.

• Increased:

• Tissue necrosis, particularly those
  involving acute injury to heart, red cells,
  kidney, skeletal muscle, liver, lung
• In all these Pt where CVS disease – suspected
• Signs – cyanosis, clubbing, peripheral edema
• H/O : Palpitations, dizziness

•   Lab inv : cholesterol > 240mg/dl
7. Serum uric ACID
• Range : 4 mg /dl to 8.5 mg/dl - Males
•         2.8 to 7.5 mg/dl – females
• Metabolic end product – nucleoprotein metabolism –
  derived purine mol
URINE ANALYSIS
The Function of Urinary System
A)  Excretion & Elimination:
           removal of organic wastes
         products from body fluids
         (urea, creatinine, uric acid)


B)  Homeostatic regulation:
         Water -Salt Balance
         Acid - base Balance

C)  Enocrine function:
          Hormones
Indication
• Disease of kidney & UI

• As a screening procedure in systemic Disease –
  diabetes , Jaundice
• Diagnosis – metabolic diseases , enteric fever

• Hormonal studies – pregnancy, steroid metabolites &
  catecholamine's
URINANALYSIS
Normal constituents
• Water – 95%
• Organic – Urea, uric Acid,
  Creatinine
• Inorganic – NaCl,
  sulphates & phosphates
• Pigments – derived bile
  pigments – colour
Divided into 3 categories
1. general physical characteristics
    & measurements
•   Appearance - clear : white &
    cloudy,
•   Colour – straw , yellow,amber
•   Odour – ammonia
•   Quantity – 1500ml
•   Specific gravity – 1.010 to
    1.030
2. Chemical Examination
•   Reaction – ph acidic 4.5-8
•   Protein – albumin
•   Glucose - no
•   Ketone – no :
•   Bilirubin - no
•   Blood - no
•   Nitrate - no
•   Uribilinogen – small amts
•   Special tests -
3. Microscopic examination of centrifuged
                               sediment

•   Cells

•   Casts

•   Bacteria

•   Parasites & yeasts

•   Spermatozoa

•   Crystals

•   Artifacts & contaminants
TYPE           Presence in         Possible causes of abnormal
               normal urine        amts of cells in urine

RBC’s          0-5 cells / hpf     Inflammatory diseses
                                   Acute glomerulonephritis
                                   Hypertension, renal infarction,
                                   trauma, stones, bleeding
                                   diseases, Use of anti
                                   coagulants


WBC’s          0-8 cells /hpf      Polynephritis, cystitis,
                                   urethritis, prostatitis,
                                   Transplant rejection,
Sq epi cells   Often present       Vaginal contamination

Tumor cells    Not often present   Tumors of
                                   Renal pelvis, Renal
                                   parenchyma,
                                   Ureters, Bladder
Casts
Urinary casts are tiny tube-shaped particles made up of white blood cells, red blood
cells, or kidney cells.
form in kidney structures called tubules. Casts are held together by a protein
released by the kidney.
Type                          Description                      causes
Hyaline casts                 Colourless,Transperent, Low RI   Strenous ex, acute
                                                               glomerulonephritis, Acute
                                                               polynephritis, Malignant
                                                               hypertension, Chronic renal
                                                               disease

Red blood cells casts         Red cells in hyaline matrix      AGN, Lupus N, Collagen
                              Yellow orange colour             disease, Renal infarction,
                              High - RI                        Malignant hypertension


Granular casts                Opaque granules in matrix        Nephrotic syn
                                                               Congestive heart failure, Acute/
                                                               chronic renal disease


Fatty casts                   Fat globules                     Nephrotic synd, DM, Mercury &
                                                               ethylene glycol poisioning


Epithelial cells              Hayaline matrix                  Glomerulonephritis, Vascuclar
                              High -RI                         disease, Toxin, Virus
Red blood cell cast in urine
                                  White blood cell cast in
                                           urine




                               Urinary casts. (A) Hyaline cast
                               (200 X); (B) erythrocyte cast
                               (100 X); (C) leukocyte cast
                               (100 X); (D) granular cast (100
                               X)
• Crystals




         Urinary crystals. (A) Calcium oxalate crystals; (B) uric acid
         crystals (C) triple phosphate crystals with amorphous
         phosphates ; (D) cystine crystals.
SALIVARY CHEMISTRY
Salivary analysis
• protective fluid - oral cavity

• Like - blood & urine- composition may alter indicating
  presence of disease
• ‘lie-detection test’

• Surrogate markers - local & systemic diseases and
  conditions.
• so-called ‘salivary biomarkers’
Application of salivary analysis
• Advancing technologies - sensitive enzyme-linked
  immunosorbent assays
• helps saliva – analyzed- study microbes, chemicals and
  immunological markers.
Advantages of salivary
            analysis
• convenient, non-invasive & painless.


• more cost-effective - testing blood assessment
  of biologically active compounds - cellular level


• blood analysis - done on compounds – protein –
  bound.
Collection of saliva samples
• Sample expectorated whole saliva
• Mixture – major & minor salivary glands + gingival
  crevicular fluid
• Collected - 1.5 to 2 hours : after meal / after overnight
  fast
Resting flow rate - whole saliva : 0.3 to 0.4 ml/minute
     flow stimulated by paraffin chewing : 1–2 ml/minute.

                                           Unstimulated/resting
      Stimulated saliva                          saliva
•   stimulated prior collection        •   collected into graduated tube or
                                           pre-weighed vial by passive
•   achieved - chew on softened
                                           drooling
    paraffin wax / washed rubber       •   flow rate per unit time -
    bands.                                 measured.
•   Topical application of 2% citric   •   cotton swabs,cotton rolls; gauze

    acid - directly to the tongue          or filter paper strips (non-
                                           volumetric collection)
                                       •   aspirated directly - floor of
                                           mouth - plastic pipette or into a
                                           graduated tube
Diagnostic Applications of
        Salivary
        Analysis
3. Infectious diseases
4. Oncology
  •   Saliva - analyzed - presence tumor markers, mutated genes
  •   exfoliated cells - saliva - used :simple, non-invasive method for
      obtaining DNA for gene analysis.
  •   diagnosis / detection - cancers distant oral cavity.

p53                     levels of salivary antibodies to p53.
CA 125                  Elevated – ovarian carcinomas
c-erbB-2 & Epidermal    Elevated – breast cancers
growth factor
Albumin                 Stomatitis associated chemotherapy
Ingested nitrate        salivary nitrate - indicative ingested nitrate
                         converted to nitrite and nitrosamine
                        contribute - development of oral and gastric carcinomas

Recurrent oral cancer   Salivary concentrations - carbohydrate antigens (Cyfra
                        21-1, TPS and CA 125) : found to significantly increase
HISTOPATHOLOGY, CYTOLOGY
           AND
   IMMUNOFLUORESCENT
         STUDIES
• Study - microscopic anatomy of cells and tissues -
  plants and animals.
• Performed by examining cells and tissues - sectioning
  and staining
• Followed by examination - light microscope or electron
  microscope
Commonly used methods
Exfoliative cytology :
• chair side
• Screening large areas, un limited repetition
• Early detection – maliganancy
• Lesions – herpes & candida – scrapping
• determining site of biopsy

Aspiration Cytology :
• Microinvasive procedure

Biopsy :
• gross & microscopic exam.
• Tissues of cells – removed living patients
BIOPSY
              USES                       COMPLICATION
•   Diagnosis of pathological     •   Hemorrhage
    lesion                        •   infection
•   Grading of tumor              •   Poor wound healing
•   Neoplastic & non neoplastic   •   Spread of tumor cells
    lesions                       •   Injury adjacent cells
•   Metastatic lesions            •   Reaction - LA
•   Evaluation of recurrence
•   Therapeutic assessment
•   Differentiation – Benign &
    malignant
Immufluorescence studies
•   Technique – antibodies or antigens are labeled with fluorescent
    dyes
•   Used - visualize - subcellular distribution of biomolecules.

•   Immunofluorescent labeled tissue sections or cultures

•   studied using a fluorescence microscope or by confocal microscopy

•   Three types of fluorescent antibody

•   Procedure - direct immunofluorescence, indirect
    immunofluorescence
Direct immunofluorescence
technique

                                           Indirect Immunofluorescence technique



  •   Auto antibodies bound to patient’s    •   detecting antibodies circulating -
      tissue                                    blood.

  •   Can be detected.                      •   monkeys esophagus - patient’s
  •   Frozen section - patient’s tissue     •   serum is added.
  •   Antihuman antibodies tagged with      •   excess serum - washed away.
      fluorescein dye is added.             •   Antihuman antibodies tagged -
  •   Excess suspension - washed                fluorescein dye added.
      away.                                 •   washed
  •   Section is viewed under               •   viewed under microscope (UV)
      microscope (uv light).
ASPIRATE
DISEASE                            Aspirate
OKC                                Thick , cheesy ,yellow granular fluid
                                   Keratin dough like consistency

Ameloblastoma                      Clear brownish yellow colour fluid
Odontogenic cyst                   Straw colored fluid : + cholesterol
                                   crystals
Sebaceous cyst                     Sebum – homogeneous & yellowish
                                   cheesy sub

Thyroglossal duct cyst             Dark amber col.
Hemangioma, varicosities, hematoma Blue blood
Aneurysm & arteriovenous fistula   Brighter red blood
Actinomycosis                      Pus with yellow granule - sulphur
Features                      Disease
Ruston bodies                 Dentigerous cyst
Reed sternberg cell           Hodgkins disease
Saw tooth appearance          Lichen planus
Picket fence / tombstone      Primodial cyst

Lipschtz bodies               Herpes simplex inf
Antischkow cell               Apthous ulcer, sickle cell, megaloblastic and fe def
                              anemia
Liesegang ring                CEOT
Cart wheel / checker board    MM
app
Lava flowing around boulder   Dentin Dysplasia

Honey comb/ swiss cheese      Adenoid cystic carcinoma of salivary gland
pattern
Cell in cells                 Hereditary benign intraepithelial keratosis
SKIN LESIONS :
LP                          Saw tooth rete pegs
Pemphigus                   Tzank cells’
Pemphigoid                  Sub epithelial vesicles - acantholysis



CYSTS
OKC                         Tomb stone basal cells & satellite cysts
DENTIGEROUS CYST            Cholesterol clefts



INFECTIONS
TB                          Langhans gaint cells & epitheloid cells
ACTINOMYCOSIS               Col of fungus- ray fungs


BONE LESIONS
FD                          Trabaculae- chinese lettern pattern

PAGETS DISEASE              Jig saw puzzle/ mosaic pattern



BENIGN & MALIGNANT TUMORS

SCC                         Multiple keratin pearl formation

Fibrosarcoma                Cells – herring bone pattern
MICROBIOLOGY
• Study - microscopic organisms

• Microbiology is a broad term which
  includes virology, mycology, parasitology, bacteriology,
  immunology and other branches.
Haematoxylin – Eosin Stain :                    Nuclei – blue black
                                                Cytoplasm – Pink or varying grades
                                                Collagen Fibers- Pink to red
                                                Muscle fibers – deep pink to red
                                                Fibrin – deep pink

Periodic Acid Solution / Periodic Acid schiff   Carbohydrate – Magenta
                                                Nucleus – Blue
                                                Glycogen – Magenta

Carbol Fuschin & distilled water                Gram + ve organisms : Violet (basophilic)
                                                Gram –ve organisms : Pink Eosinophilic



Acid Fast stain ( Ziehl Neelsen’s stain)        Mycobactterium tuberculi & Mycobacterium
                                                leprae
                                                { red} tissue & other organisms – blue


 Mycobacterium retains stain after decolorization – thick lipid mycolic acid
SEROLOGY AND
IMMUNOHISTOCHEMISTRY
METHODS OF DETECTION OF ANTIBODIES

 1. Immuno-precipitation Assays
    = detect antibodies in solution
    = qualitative indication of the presence of
    antibodies
    = end-point is visual flocculation of the antigen and
    antibody in suspension

 2. Complement Fixation
     = based on the activation or fixation of
     complement following binding of complement
     factors to Ag-Ab immune complexes
3. Neutralization
    = effectively of an organism or activity of toxin is neutralized by
            specific antibody
    = rarely used for diagnostic purposes
    = mainly used to detect antibody formation after vaccination


4. Particle Agglutination

    = relatively simple and fast
    = capable of detecting lower concentration of antibodies
    = designed to detect antibodies to viruses, subsequent to
    interaction or vaccination
    = utilize Ag coated latex particles, coal particles,
    = direct and indirect methods
5. Immunofluorescence
–     requires use of microscope equipped      provide ultraviolet
     illumination or

–     an instrument capable of irradiating the assay with UV light and
     detecting resultant fluorescence with a fluorometer

6. Enzyme Immunoassay
–     most sensitive
–    usually indirect assay that depends on the use of an antihuman
     IgG or IgM antibody conjugate

–     antibody conjugate (if present) is made to attach to enzyme which
     catalyzes conversion of substrate to a colored product which
             will then be read with the use of a spectrophotometer

7. Radioimmunoassay
     = high sensitivity
Tests for HIV
Laboratory diagnosis of HIV infection
❑ Detection of anti-HIV antibodies
❑ Detection of antigen
❑ Detection of viral nucleic acid
❑ Virus isolation.
Investigations for HIV non-specific tests
Lymphocytopenia below 2000 cu.mm
Decrease in CD4 count
Low t4:t8 ratio (t helper/t suppressor cell ratio) increase in
IgG and IgA.
Specific Tests
Primary tests
❑ EIA/ELISA
❑ Polymerase chain reaction

Confirmatory tests
❑ Western blot
❑ RIPA
❑ Immunofluorescence assay
❑ DNA/RNA amplification tests
PCR – Polymerase Chain Reaction –
• uses - amplify viral RNA from blood,
• detect even small amounts of virus in newly infected person.
• expensive, time consuming, and not readily available
RIPA –
• done when antibody levels are low or Western Blot results -
   unclear
• expensive, difficult to perform, and not often used.
Immunofluorescence Assay –
• Confirmatory test used when Western Blot results are unclear.
• used instead of a Western Blot after an ELISA test.
 DNA/RNA Amplification Tests –
• Tests similar to PCR
• used when the result of a western blot is unclear.
Radiological
• Radiographs – recomended for demonstrating varios
  lesións
Region required   Standard views              Additional views

SKULL             PA, Lateral skull           SMV

FACIAL BONES      OM,Lateral                  Red exposure SMV

PNS               OM for max antrum           Upper occlusal/lat
                                              SMV.OPG,CT

Orthodontics      OPG, Cephalometric
                  skull

Nasal bones       OM – 30 ,Lateral,

MANDIBLE          OPG                         Lat oblique, PA mandible
                                              MAnd.Occlusal

TMJ               Transcranial,Lat oblique,   Transpharyngeal,Arthrog
                  OPG                         raphy, reberse townes,
                                              Ct,,MRI
SPECIAL RADIOGRAPHIC TECNIQUES

•   CT
•   Digital Radiography
•   Subtraction radiography
•   Radionuclide imaging
•   Sonography
Conclusion
THANK YOU
TESTS FOR SPECIFIC
    DISEASES
Tests for Syphilis
• Diagnosis - achieved either by direct identification -
  pathogen - serological findings & treponema pallidum
Serology




•   confirmatory test - FTAABS - golden standard
•   enzyme immunoassays, Western blot technique
•   Serology in congenital syphilis: Finding specifi c IgM anti-
    treponemal antibodies is helpful in diagnosing congenital infection
•   If titers > in infant then mother
 laboratory  investigations

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laboratory investigations

  • 2. Contents • Introduction • Tests done by dentists • Hematological • Urine Analysis • Biochemistry :Renal function tests Tests – diseases of bone Liver function tests Lipid Analysis • Immunological investigations • Histopathology, cytology and immunofluorescent studies Microbiology • Radiological • Conclusion • References
  • 3. INTRODUCTION • Diagnosis & identification - disease by careful investigation o patients signs, symptoms and history • Carefully crafted history and physical examination - satisfactory – diagnosis • Times when more information is required through the use of diagnostic tests. • Clinical and/or lab data must be used to distinguish between different diagnoses.
  • 4. importance of laboratory investigations • laboratory tests - important in assisting & management of the patient during treatment of disease besides diagnosis. • Used- 1. screen - disease in asymptomatic individual 2. to establish or exclude presence of diseases in symptomatic patients 3. assist the practitioner in the management of the patient.
  • 5. important characteristics of a laboratory test • Accuracy • Cost • Interfering factors • Precision • Reference range • Sensitivity • Specificity
  • 6. TESTS ADVISED BY DENTISTS Acc. Sonis, Fazio & Fang
  • 7. Depending upon the specimens submitted 1. Hematological 2. Biochemical 3. Immunological 4. Histopathological – histological & Immunofluorescent 5. Microbial 6. Radiological
  • 8. Depending upon the organs involved • Heart • Skin • Vascular system • Eye • Respiratory system • Bones & joints • GIT • Skeletal muscle • Liver, gallbladder & pancreas • CVS • Kidney & genitourinary • Mental illness • Female genital • Endocrine system • Breast • Infectious diseases • Immunological diseases
  • 9. Laboratory investigation  Biochemistry :  Urine analysis  Salivary chemistry  Histopathology, cytology, Immunofluorescent studies  Serology & immunohistochemistry  Radiology
  • 10. BLOOD CHEMISTRY TESTS A. . Estimation of electrolytes B. . Renal function C. .Liver function D. .Thyroid Function E. . Metabolic bone disease F. . Other tests .
  • 11. A. Estimation of electrolytes • Maintain fluid levels inside & outside cells • Osmotic gradient – nerve condtn. Muscle func,hydration & maintaining Ph levels 1. SODIUM : • Loss of sodium results extracellular fluid volume • Circulation, renal function, and central nervous system function. • Normal : 136-145 m Eq/l
  • 12. Feel weak, confusion, paralysis, sezuires Decrease consciousness, vomiting, fatigue, muscle weakness
  • 13. 2. Chloride • Normal : 95 to 108 mEq/l. • Important - maintenance of acid-base balance
  • 14. 3. Potassium • determines neuromuscular irritability. • or conc. impair - ability of muscle tissue contract • Normal - 3.5 to 5.2 mEq/l.
  • 15. 4. Bicarbonate • Bicarbonate–carbonic acid buffer - maintaining normal pH of body fluids. • basis for assessing acid-base balance. • Normal : 24 to 30 mEq/l.
  • 16. Dentists • Electrolytes - • Maintain - Ph • Proteins are denatured and digested • Enzymes lose their ability to function and death may occur. • possibly life threatening. • could require hospitalization and intravenous electrolytes.
  • 17. b. renal function test 1. blood urea nitrogen • Normal value of BUN in adults: 8 – 18 mg %. 10 – 20 mg/dl sonis symptom - >50 mg/dl
  • 18. Clinical significance : • Pre renal causes – reduced flow to kidney, shock, blood loss, dehydration, increased protein catabolism, injuries, burns, fever • Renal causes – acute renal failure, glomerulonephritis, malignant hypertension, chronic renal failure, DM • Post renal causes – urinary obstruction by stones, tumors
  • 19. Decreased blood urea nitrogen – • Poor nutrition • High fluid intake • Excessive administration of i.v fluids • Liver damage • Late pregnancy, infancy, acromegaly.
  • 20. 2. creatinine • Creatinine - catabolic product of creatinine phosphate - used for skeletal muscle contraction. • excreted entirely - kidneys • amount in blood - directly proportional to renal excretory function • diagnose impaired renal function • Normal range – 0.5 – 1.6mg% or 1.5 mg/dl • symptom > 4mg/dl
  • 21. INCREASED CREATININE LEVELS: • Kidney disease • Decreased: Pregnancy – NS • decreased – improve function
  • 22. Dentists • O/E : Pale, fluid retention – edema, Inc IJP, asitics' - advice Investigations • Anemic, Acidosis – K+ retention • BT prolonged • Rx – avoid anti platelet drugs – Aspirin NSAIDs, nephrotoxic drugs • Consent – physician
  • 23. C. LIVER FUNCTION Serum bilirubin Urine urobilinogen Serum albumin – globulin Serum alkaline phosphatase AST & ALT
  • 24. 1.serum bilirubin • Bilirubin : excreted - intestines. • Intestines converted - colorless compound - urobilinogen. • Level of bilirubin elevates - excessive hemolysis • Inability of liver to eliminate bilirubin (liver damage or obstruction of bile duct) • yellow color - product • skin, mucous membranes, sclera of the eye, plasma and urine • Excess bilirubin - associated - hepatitis
  • 25. • Jaundice - total serum bilirubin rises • Normal Value → 0.1 to 1.2 mg / 100 ml 5.1–17.0 micro mol/L High values: • Hemolytic Anemia • Biliary obstruction • Hepatitis • Malignant Hepatic Disease
  • 26. 2. SERUM GLUTAMIC OXALOACETIC TRANSAMINASE (SGOT) SERUM GLUTAMIC PYRUVIC TRANSAMINASE (SGPT) • enzymes -large amounts - liver, heart, kidney and skeletal muscle • metabolism of amino acids and carbohydrates. • NORMAL VALUES : • SGOT also called Aspartate Aminotransferase 6 -25 I U/ L • SGPT also called Alanine Aminotransferase 3– 26 I U/ L
  • 27. • SGOT -sensitive indicator - myocardial necrosis, significant rises occurs • Hepatitis & other liver disease + associated liver necrosis- SGOT and SGPT
  • 28. screening for hepatitis b &c • Detection of hepatitis B surface antigen: • Hepatitis B virus (HBV) is a double stranded DNA spherical particle with a double shell. - referred as the Dane particle. • Antigens which have so far been associated with HBV are as follows ; • Hepatitis B surface antigen (HBsAg) : This was first noticed in the blood of an Australian aborigine hence earlier was termed as Australia antigen. • Hepatitis B core antigen (HBcAg) • Hepatitis B e antigen (HBeAg).
  • 29. • Infection - HBV i- entry of virus through the skin or mucous membranes into the blood or body fluids. Routes of transmission • transfusion of blood or blood products. • HBV contaminated needles, syringes or prickers. • Close personal contact with a person with hepatitis B • Contact with blood specimen (Laboratory personnel those who handle the specimen) • Transmission by blood sucking insects or bed bugs.
  • 31. • detection of hepatitis – b surface antigen (hbsag) by direct enZyme linKed immunosorbent assay (elisa) : • HBsAg formerly called Australia antigen is • Reduce incidence of post-transfusion hepatitis, testing for HbsAg is mandatory for blood products intended for human use. • Care – Rx – viral particles – saliva & other body secretions.
  • 32. • Hepatitis A – SGOT, SGPT – • Hepatitis B – SGOT,SGPT & LDH – • Hepatitis C – SGOPT & SGPT • Dentists – possible signs – icterus, fever ../ H/O • Lab inv - • Viral hepatitis – infectious B • Exposed inf blood & saliva- Precautions
  • 33. • Signs • H/O: Alcohol abuse, hepatitis ,abnormal bleeding • Features – improper function Liver – Def production clotting factors – PT elevated • So investigation & physicians opinion – needed – precautions
  • 34. 3. serum albumin and globulin • total protein • important - coagulation, transport hormones, act on antibodies • act - buffers with enzymes • help - maintain osmotic pressure.
  • 35. • Serum albumin - synthesized in liver • globulins - produced - plasma cell • Normal Values: • Total Protein 6.0 to 7.8 g / 100 ml or 6.0 to 8.3 gm/dL • Albumin 3.2 to 5.6 g / 100 ml • Globulin 2.3 to 3.5 g / 100 ml • A / G Ratio 1.5 to 1 to 2.5:1
  • 36. Total proteins High Albumin - rare but dehydration and shock. Low Albumin : same as proteins. High Globulin : Multiple myeloma, Nephrosis, Chronic infections, Collagen diseases, Liver diseases Low Globulin : Burns and severe malnutrition.
  • 37. 5.Alkaline phosphatase • high concentration - growing bone, bile, placenta. • Normal level: 30 to 115 U/l. Increased A. In children B. Osteoblastic bone disease C. Hepatic disease / bileobstruction - stone, stricture, neoplasm. D. Pregnancy. Decreased: • Hypophosphatasia, hypothyroidism, malnutrition.
  • 38. Normal values and changes in two types of Jaundice Hepatocellular Uncomplicated Tests Normal Values Jaundice Obstructive Jaundice Bilirubin - Direct 1. – 0.3 mg/dL Increased Increased -Indirect 2. – 0.7 mg/dL Increased Increased Urine bilirubin None Increased Increased Serum albumin/Total Albumin, 3.5-5.5 g/dL Albumin decreased Unchanged protein Total protein, 6.5-8.4 g/dL Alkaline phosphatase 30-115 units /L Increased (+) Increased (++++) Prothrombin time INR of 1.0 – 1.4 After Prolonged if damage Prolonged if obstruction Vitamin K, 10% increase severe and does not marked, but responds to in 24 hours respond to parenteral parenteral vitamin K Vit K ALT, AST ALT, 5-35 units/L, AST, 5-40 Increased in Minimally increased units/L hepatocellular damage, viral hepatitis
  • 40. Reference values of thyroid function test Test Range TSH 0.5 - 4.7mU/L T3 0.92-2.78nmol/L T4 58-140 nmol/L FT4 10.3-35pmol/L
  • 41.
  • 42. HYPO THYROIDISM HYPER THYROIDISM • Catecholamines in LA + • CNS depression – present stress in dental operatory • Administration of narcotic • Ppt thyroid Astorm analgesics increase risk • Characterised High fever, CNS depression& collapse psycosis, CNS depresssion, vomiting diaorrhea – risk CHF Pt asked for Heat & cold intolerance, wt gain /loss, change in appetite bowel habits , muscle weakness and palpitations
  • 43. E. Metabolic Bone Disease • Jaw lesions – radiographic examination • Systemic jaw diseases- • Pagets disease, FD, Primary & Sec. Hyperparathyroidism, Osteoporosis, MM, Osteogenic sarcoma or metastatic malignancy • Serum Ca, P,& alkaline phosphatase -
  • 44. Serum Ca, P • Serum cal I / serum phosphorus • Measured : mg/dl • serum Ca conc. X serum P conc. = constant • 30 to 40 mg/dl = Normal adults • 50 to 60 mg/ dl = growing children
  • 45. GA & surgical procedures – cardiac arrythmmias, heart block • Hypercalcaemia - excessive skeletal calcium release, increased intestinal calcium absorption, or decreased renal calcium excretion. • serum Ca – hypo proteinmia, due decreased binding by serum protein & renal disease.
  • 46. 2. Alkaline phosphatase • Oestoblasts • enzyme - produced - small amounts –liver • larger amounts by osteoblasts. • active in bone formation and therefore is found at higher levels there. • result increased osteoblastic activity • In association – obstructive liver disease, amyloid disease, leukemia & sarcoidosis
  • 47. Normal values for serum Alkaline phosphatase • King Armstrong Units – 4 to 13 • Brodansky Units - 1.5 to 4.5 • International Units - 30 to 85 IU
  • 48. High Alkaline Phosphatase values : • Obstructive liver disease • Metastatic carcinoma involving bones • Hyper parathyroidism • Pagets Disease of Bone • Osteo malacia • Rickets • Amyloid Disease • Leukemia • Sarcoidosis Low Alkaline Phosphatase : • Hypophosphatasia • Hypothyroidism • Scurvy
  • 49. Values Serum calcium Serum Serum phosphates mg/dl phosphorus Units /dl P/dl Normal 8.8 to 10.5 2 to 5 1-4 Rickets Normal Decreased – Increased 20 to 40 x Exc tetany normal Osteomalacia Decreased Decreased Little if any change Paget’s Disease Normal Normal Occasionally elevated Hyperparathyroidism Increased Decreased Increased 2 to 50 x normal Osteogenic normal Normal Slightly increase Imperfecta Solitary Bone cysts normal normal normal Metatstatic Oseous May be elevated Normal Normal / slightly Disorders elevated Tetany 7 mg ca/dl or less Normal / elevated Normal
  • 50.
  • 51. G. Other Tests 1. Acid phosphatase • Phosphatases active at pH 4.9 are present - high conc. • prostate gland, erythrocytes, platelets, reticuloendothelial cells, liver, spleen, and kidney. • Normal levels: 0.8 IU/l. • Increased: carcinoma of the prostate
  • 52. 2. Serum amylase • Normal level: 5 to 75 IU/l. • Increased: • Acute pancreatitis, pseudocyst of the pancreas, • obstruction of pancreatic ducts (carcinoma, stone, stricture,duct sphincter spasm after morphine), and parotitis. • Decreased: • Acute and chronic hepatitis • Pancreatic insufficiency, • toxemia of pregnancy.
  • 53. 3. Serum lipase Normal level: 0.2 to 1.5 units. Increased: • acute or exacerbated pancreatitis • obstruction of pancreatic ducts- stone or neoplasm.
  • 54. 4. Serum cholesterol • Normal Range : 160 to 300 mg/dl • > 300 mg/dl : dietary reduction – animal fats, eggs & foods cholesterol levels • Risk – artherosclerotic CVD • Elevated : • hypothyroidism, obese; elderly diabetic & nephrotic syndrome
  • 55.
  • 56. Check on triglycerides • HDL : good" cholesterol - removes excess cholesterol from the blood and takes it to the liver • LDL: BAD: High levels- linked - increased risk of heart and blood vessel disease, inlcuding coronary artery disease, heart attack and death.
  • 57. 5. Creatine phosphokinase (CPK) • Male: 50 to 180 IU/l, female: 50 to 60 IU/l. • Myocardial infarction, trauma to muscle, malignant • Hyperthermia muscular dystrophies, polymyositis, severe Muscular exertion (jogging), hypothyroidism
  • 58. 6. Lactate dehydrogenase (LDH) • Normal level: 45 to 100 U/l. • Increased: • Tissue necrosis, particularly those involving acute injury to heart, red cells, kidney, skeletal muscle, liver, lung
  • 59. • In all these Pt where CVS disease – suspected • Signs – cyanosis, clubbing, peripheral edema • H/O : Palpitations, dizziness • Lab inv : cholesterol > 240mg/dl
  • 60. 7. Serum uric ACID • Range : 4 mg /dl to 8.5 mg/dl - Males • 2.8 to 7.5 mg/dl – females • Metabolic end product – nucleoprotein metabolism – derived purine mol
  • 61.
  • 63. The Function of Urinary System A)  Excretion & Elimination: removal of organic wastes products from body fluids (urea, creatinine, uric acid) B)  Homeostatic regulation: Water -Salt Balance Acid - base Balance C)  Enocrine function: Hormones
  • 64. Indication • Disease of kidney & UI • As a screening procedure in systemic Disease – diabetes , Jaundice • Diagnosis – metabolic diseases , enteric fever • Hormonal studies – pregnancy, steroid metabolites & catecholamine's
  • 65. URINANALYSIS Normal constituents • Water – 95% • Organic – Urea, uric Acid, Creatinine • Inorganic – NaCl, sulphates & phosphates • Pigments – derived bile pigments – colour
  • 66. Divided into 3 categories 1. general physical characteristics & measurements • Appearance - clear : white & cloudy, • Colour – straw , yellow,amber • Odour – ammonia • Quantity – 1500ml • Specific gravity – 1.010 to 1.030
  • 67. 2. Chemical Examination • Reaction – ph acidic 4.5-8 • Protein – albumin • Glucose - no • Ketone – no : • Bilirubin - no • Blood - no • Nitrate - no • Uribilinogen – small amts • Special tests -
  • 68. 3. Microscopic examination of centrifuged sediment • Cells • Casts • Bacteria • Parasites & yeasts • Spermatozoa • Crystals • Artifacts & contaminants
  • 69. TYPE Presence in Possible causes of abnormal normal urine amts of cells in urine RBC’s 0-5 cells / hpf Inflammatory diseses Acute glomerulonephritis Hypertension, renal infarction, trauma, stones, bleeding diseases, Use of anti coagulants WBC’s 0-8 cells /hpf Polynephritis, cystitis, urethritis, prostatitis, Transplant rejection, Sq epi cells Often present Vaginal contamination Tumor cells Not often present Tumors of Renal pelvis, Renal parenchyma, Ureters, Bladder
  • 70. Casts Urinary casts are tiny tube-shaped particles made up of white blood cells, red blood cells, or kidney cells. form in kidney structures called tubules. Casts are held together by a protein released by the kidney. Type Description causes Hyaline casts Colourless,Transperent, Low RI Strenous ex, acute glomerulonephritis, Acute polynephritis, Malignant hypertension, Chronic renal disease Red blood cells casts Red cells in hyaline matrix AGN, Lupus N, Collagen Yellow orange colour disease, Renal infarction, High - RI Malignant hypertension Granular casts Opaque granules in matrix Nephrotic syn Congestive heart failure, Acute/ chronic renal disease Fatty casts Fat globules Nephrotic synd, DM, Mercury & ethylene glycol poisioning Epithelial cells Hayaline matrix Glomerulonephritis, Vascuclar High -RI disease, Toxin, Virus
  • 71. Red blood cell cast in urine White blood cell cast in urine Urinary casts. (A) Hyaline cast (200 X); (B) erythrocyte cast (100 X); (C) leukocyte cast (100 X); (D) granular cast (100 X)
  • 72. • Crystals Urinary crystals. (A) Calcium oxalate crystals; (B) uric acid crystals (C) triple phosphate crystals with amorphous phosphates ; (D) cystine crystals.
  • 74. Salivary analysis • protective fluid - oral cavity • Like - blood & urine- composition may alter indicating presence of disease • ‘lie-detection test’ • Surrogate markers - local & systemic diseases and conditions. • so-called ‘salivary biomarkers’
  • 75. Application of salivary analysis • Advancing technologies - sensitive enzyme-linked immunosorbent assays • helps saliva – analyzed- study microbes, chemicals and immunological markers.
  • 76. Advantages of salivary analysis • convenient, non-invasive & painless. • more cost-effective - testing blood assessment of biologically active compounds - cellular level • blood analysis - done on compounds – protein – bound.
  • 77. Collection of saliva samples • Sample expectorated whole saliva • Mixture – major & minor salivary glands + gingival crevicular fluid • Collected - 1.5 to 2 hours : after meal / after overnight fast
  • 78. Resting flow rate - whole saliva : 0.3 to 0.4 ml/minute flow stimulated by paraffin chewing : 1–2 ml/minute. Unstimulated/resting Stimulated saliva saliva • stimulated prior collection • collected into graduated tube or pre-weighed vial by passive • achieved - chew on softened drooling paraffin wax / washed rubber • flow rate per unit time - bands. measured. • Topical application of 2% citric • cotton swabs,cotton rolls; gauze acid - directly to the tongue or filter paper strips (non- volumetric collection) • aspirated directly - floor of mouth - plastic pipette or into a graduated tube
  • 79. Diagnostic Applications of Salivary Analysis
  • 80.
  • 82. 4. Oncology • Saliva - analyzed - presence tumor markers, mutated genes • exfoliated cells - saliva - used :simple, non-invasive method for obtaining DNA for gene analysis. • diagnosis / detection - cancers distant oral cavity. p53 levels of salivary antibodies to p53. CA 125 Elevated – ovarian carcinomas c-erbB-2 & Epidermal Elevated – breast cancers growth factor Albumin Stomatitis associated chemotherapy Ingested nitrate salivary nitrate - indicative ingested nitrate converted to nitrite and nitrosamine contribute - development of oral and gastric carcinomas Recurrent oral cancer Salivary concentrations - carbohydrate antigens (Cyfra 21-1, TPS and CA 125) : found to significantly increase
  • 83.
  • 84. HISTOPATHOLOGY, CYTOLOGY AND IMMUNOFLUORESCENT STUDIES
  • 85. • Study - microscopic anatomy of cells and tissues - plants and animals. • Performed by examining cells and tissues - sectioning and staining • Followed by examination - light microscope or electron microscope
  • 86. Commonly used methods Exfoliative cytology : • chair side • Screening large areas, un limited repetition • Early detection – maliganancy • Lesions – herpes & candida – scrapping • determining site of biopsy Aspiration Cytology : • Microinvasive procedure Biopsy : • gross & microscopic exam. • Tissues of cells – removed living patients
  • 87. BIOPSY USES COMPLICATION • Diagnosis of pathological • Hemorrhage lesion • infection • Grading of tumor • Poor wound healing • Neoplastic & non neoplastic • Spread of tumor cells lesions • Injury adjacent cells • Metastatic lesions • Reaction - LA • Evaluation of recurrence • Therapeutic assessment • Differentiation – Benign & malignant
  • 88. Immufluorescence studies • Technique – antibodies or antigens are labeled with fluorescent dyes • Used - visualize - subcellular distribution of biomolecules. • Immunofluorescent labeled tissue sections or cultures • studied using a fluorescence microscope or by confocal microscopy • Three types of fluorescent antibody • Procedure - direct immunofluorescence, indirect immunofluorescence
  • 89. Direct immunofluorescence technique Indirect Immunofluorescence technique • Auto antibodies bound to patient’s • detecting antibodies circulating - tissue blood. • Can be detected. • monkeys esophagus - patient’s • Frozen section - patient’s tissue • serum is added. • Antihuman antibodies tagged with • excess serum - washed away. fluorescein dye is added. • Antihuman antibodies tagged - • Excess suspension - washed fluorescein dye added. away. • washed • Section is viewed under • viewed under microscope (UV) microscope (uv light).
  • 90.
  • 91. ASPIRATE DISEASE Aspirate OKC Thick , cheesy ,yellow granular fluid Keratin dough like consistency Ameloblastoma Clear brownish yellow colour fluid Odontogenic cyst Straw colored fluid : + cholesterol crystals Sebaceous cyst Sebum – homogeneous & yellowish cheesy sub Thyroglossal duct cyst Dark amber col. Hemangioma, varicosities, hematoma Blue blood Aneurysm & arteriovenous fistula Brighter red blood Actinomycosis Pus with yellow granule - sulphur
  • 92. Features Disease Ruston bodies Dentigerous cyst Reed sternberg cell Hodgkins disease Saw tooth appearance Lichen planus Picket fence / tombstone Primodial cyst Lipschtz bodies Herpes simplex inf Antischkow cell Apthous ulcer, sickle cell, megaloblastic and fe def anemia Liesegang ring CEOT Cart wheel / checker board MM app Lava flowing around boulder Dentin Dysplasia Honey comb/ swiss cheese Adenoid cystic carcinoma of salivary gland pattern Cell in cells Hereditary benign intraepithelial keratosis
  • 93. SKIN LESIONS : LP Saw tooth rete pegs Pemphigus Tzank cells’ Pemphigoid Sub epithelial vesicles - acantholysis CYSTS OKC Tomb stone basal cells & satellite cysts DENTIGEROUS CYST Cholesterol clefts INFECTIONS TB Langhans gaint cells & epitheloid cells ACTINOMYCOSIS Col of fungus- ray fungs BONE LESIONS FD Trabaculae- chinese lettern pattern PAGETS DISEASE Jig saw puzzle/ mosaic pattern BENIGN & MALIGNANT TUMORS SCC Multiple keratin pearl formation Fibrosarcoma Cells – herring bone pattern
  • 95. • Study - microscopic organisms • Microbiology is a broad term which includes virology, mycology, parasitology, bacteriology, immunology and other branches.
  • 96. Haematoxylin – Eosin Stain : Nuclei – blue black Cytoplasm – Pink or varying grades Collagen Fibers- Pink to red Muscle fibers – deep pink to red Fibrin – deep pink Periodic Acid Solution / Periodic Acid schiff Carbohydrate – Magenta Nucleus – Blue Glycogen – Magenta Carbol Fuschin & distilled water Gram + ve organisms : Violet (basophilic) Gram –ve organisms : Pink Eosinophilic Acid Fast stain ( Ziehl Neelsen’s stain) Mycobactterium tuberculi & Mycobacterium leprae { red} tissue & other organisms – blue Mycobacterium retains stain after decolorization – thick lipid mycolic acid
  • 97.
  • 99.
  • 100. METHODS OF DETECTION OF ANTIBODIES 1. Immuno-precipitation Assays = detect antibodies in solution = qualitative indication of the presence of antibodies = end-point is visual flocculation of the antigen and antibody in suspension 2. Complement Fixation = based on the activation or fixation of complement following binding of complement factors to Ag-Ab immune complexes
  • 101. 3. Neutralization = effectively of an organism or activity of toxin is neutralized by specific antibody = rarely used for diagnostic purposes = mainly used to detect antibody formation after vaccination 4. Particle Agglutination = relatively simple and fast = capable of detecting lower concentration of antibodies = designed to detect antibodies to viruses, subsequent to interaction or vaccination = utilize Ag coated latex particles, coal particles, = direct and indirect methods
  • 102. 5. Immunofluorescence – requires use of microscope equipped provide ultraviolet illumination or – an instrument capable of irradiating the assay with UV light and detecting resultant fluorescence with a fluorometer 6. Enzyme Immunoassay – most sensitive – usually indirect assay that depends on the use of an antihuman IgG or IgM antibody conjugate – antibody conjugate (if present) is made to attach to enzyme which catalyzes conversion of substrate to a colored product which will then be read with the use of a spectrophotometer 7. Radioimmunoassay = high sensitivity
  • 103. Tests for HIV Laboratory diagnosis of HIV infection ❑ Detection of anti-HIV antibodies ❑ Detection of antigen ❑ Detection of viral nucleic acid ❑ Virus isolation. Investigations for HIV non-specific tests Lymphocytopenia below 2000 cu.mm Decrease in CD4 count Low t4:t8 ratio (t helper/t suppressor cell ratio) increase in IgG and IgA.
  • 104. Specific Tests Primary tests ❑ EIA/ELISA ❑ Polymerase chain reaction Confirmatory tests ❑ Western blot ❑ RIPA ❑ Immunofluorescence assay ❑ DNA/RNA amplification tests
  • 105. PCR – Polymerase Chain Reaction – • uses - amplify viral RNA from blood, • detect even small amounts of virus in newly infected person. • expensive, time consuming, and not readily available RIPA – • done when antibody levels are low or Western Blot results - unclear • expensive, difficult to perform, and not often used. Immunofluorescence Assay – • Confirmatory test used when Western Blot results are unclear. • used instead of a Western Blot after an ELISA test. DNA/RNA Amplification Tests – • Tests similar to PCR • used when the result of a western blot is unclear.
  • 107. • Radiographs – recomended for demonstrating varios lesións
  • 108. Region required Standard views Additional views SKULL PA, Lateral skull SMV FACIAL BONES OM,Lateral Red exposure SMV PNS OM for max antrum Upper occlusal/lat SMV.OPG,CT Orthodontics OPG, Cephalometric skull Nasal bones OM – 30 ,Lateral, MANDIBLE OPG Lat oblique, PA mandible MAnd.Occlusal TMJ Transcranial,Lat oblique, Transpharyngeal,Arthrog OPG raphy, reberse townes, Ct,,MRI
  • 109. SPECIAL RADIOGRAPHIC TECNIQUES • CT • Digital Radiography • Subtraction radiography • Radionuclide imaging • Sonography
  • 112. TESTS FOR SPECIFIC DISEASES
  • 113. Tests for Syphilis • Diagnosis - achieved either by direct identification - pathogen - serological findings & treponema pallidum
  • 114. Serology • confirmatory test - FTAABS - golden standard • enzyme immunoassays, Western blot technique • Serology in congenital syphilis: Finding specifi c IgM anti- treponemal antibodies is helpful in diagnosing congenital infection • If titers > in infant then mother