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Disease Testing
Group 9
Fatima Tu Zahra
Komal Binte Ajmal
Maham Nadeem
Muhammad Ahmed
Saifullah khan
Sumayya Irfan
Autoimmune
Diseases
Ahmed
Autoimmunity
Hallmark of Autoimmunity
Self reactive T-cells
Autoantibodies
Inflammation
Laboratory Diagnosis
Cant be done by a single test
Several tests are needed to be done
Complete blood count
Comprehensive metabolic panel
Immunological studies
Serologies
Flow cytometry
Cytokine analysis
HLA typing
Disease activity
Erythrocyte sedimentation rate (ESR)
Blood analysis
Normochromic anemia
Platelet count
WBC count
Leukopenia and Thrombocytopenia common in SLE
patients
Serology
Abnormalities in serum level of specific enzymes
Abnormalities in metabolic processes
E.g. Autoimmune Hepatitis:
Elevated Transaminase
Elevated Bilirubin
Elevated serum proteins
Non-specific markers : Same in drug toxicity
Coagulation Studies
Antiphospholipid Syndrome
Prolongation of Activated Partial Thromboplastin Time (aPTT) and Prothrombin Time (PT)
Inhibition of Clotting factor
Sarcoidosis
Hypercalcemia
Autoimmune Inflammatory Myopathies
(Dermatomyositis, Polymyositis and Inclusion body myositis)
Urinalysis
Glomerulonephritis and Interstitial nephritis
Proteinuria
Hamaturia
Active sediment of RBCs and WBCs
Ferritin
Komal Binte Ajmal
Ferittin
Iron binding protein
It sequesters iron in a non toxic and
readily available form
Hyperferitinemia
Hyperferitinemia is associated with inflammation due to
infection , malignancies or due to auto immune diseases
SLE
Ferritin levels were high in the urine of patients with nephritis ,
pleural fluid and in the CSF of patients suffering from meningitis
Cont
THYRODITIS
Hormonal pathways are involved in the regulation of Ferritin
Elevated Ferritin levels
POLYMYOSTITIS /DERMATOMYOSTITIS
Inflammatory auto immune myopathy characterized by muscle weakness
and visceral movement
C reactive protein
Pentamic protein found in blood plasma
Synthesized by liver
Also an inflammatory marker
Detect inflammation which can be ether
due
to
diseases
infection
or
auto
immune
Significance Of CRP
The level of CRP in the blood is normally low.
A high or increasing amount of CRP in the blood suggests the presence of
inflammationbut will not identify its location or the cause. In individuals
suspected of having a seriousbacterialinfection, a high CRP can be
confirmatory. In people withchronicinflammatory conditions, high levels
of CRP suggest a flare-up or that treatment has not been effective.
If the CRP level is initially elevated and drops, it means that the
inflammation or infection is subsiding and/or responding to treatment.
DISEASES
RA
SLE
Vasculitis
LIMITATIONS
Non specific It only detects the presence of inflammation
but do not diagnose the cause
ESR
Commonly known as erythrocyte sedimentation rate
Simple , inexpensive but non-specific test
This test is used to measure the inflammation in body which can
either be a result of
1)infection
2)cancer
3)auto immune disorders
The test measures how fast the RBCs fall to the bottom of a
thin long tube
SIGNIFICANCE OF TEST IN
AUTO-IMMUNE DISORDERS
Elevated levels are shown which indicate inflammation
DISEASES
rheumatoid arthritis , SLE are most common
Limitations
Limitations of this test
It is a non specific test , it just indicates the presence of
inflammation but do not tell the exact location , cause and
diagnosis of disease
Inflammatory Markers
Maham Nadeem
Ceruloplasmin
Glycoprotein
Produced in liver
Carries or transports more than 95% of copper in
blood plasma
Plays a role iniron metabolism
Mechanism
Behaves as an enzyme (ferroxidase)
Catalyzes oxidation of Fe2+(ferrous iron) into Fe3+(ferric
iron)
Assists in transport in the plasma in association with
transferrin
Essential for normal movement of iron from cells to plasma
Periodontitis
A set of inflammatorydiseasesaffecting the tissues that
surround and support theteeth
Risk identified and quantified by biomarkers
Creates area of local inflammation indicated by elevated
serum levels of various pro-inflammatory markers
specially ceruloplasmin.
Inflammatory states
Pregnancy
Lymphoma
Rheumatoid arthritis
Alzheimer's disease
Haptoglobin
Protein
Encoded by theHPgene.
Produced in response to tissue injury
Mechanism
Inintravascular hemolysis, free
In extravasclar hemolysis:
Macrophages phagocytose erythrocytes
Hb not released into circulation
No Hb-haptoglobin complex formed
Serum Haptoglobin levels rise
Inflammation
Malignancy
Surgery
Trauma
Peptic ulcer disease
Ulcerative colitis
Albumin
Serum protein
Synthesized by the liver
Main function is to regulate thecolloidal osmostic
pressureof blood
Inflammation results in a lower synthesis rate
Decreased serum concentrations (negative acute
phase protein
Mechanism
Distributed between the vascular and interstitial spaces
During inflammation, IL6 increases
Results in albumin being pulled from the intravascular spaces and
circulating to the liver until theinflammatory diseaseresolves
Liver starts producing other proteins needed to fight the
inflammation.
Inflammation simultaneously decreases albumin synthesis and
increases albumin catabolism.
Autoimmune diseases
Infections
Malignancies
Trauma
Surgery, burn injuries
Crohns disease
Auto-antibodies
Saif
Auto-antibody
Auto anti-bodies are the proteins that are directed against
Types of auto-antibodies
Arthritis.
Specificity=70%
sensitivity=70%
Progressive joint morbidity and rheumatoid nodules.
Sjogren's syndrome, SLE, Cryoglobulinemia, interstitial
fibrosis and silicosis.
Diagnosis with ELISA.
Anti-citrullinated peptide
antibody.
Inflammation in RA.
Activation of peptidylarginine deiminase which incorporates
citrulline into certain proteins.
Antibody against CP.
Specificity= 95%
sensitivity= 70%.
antigens
Hallmarks the serologic diagnosis of SLE.
Also present in some non-rheumatic disorders like Hashimoto and
Graves disease.
ELISA method.
Subtypes: anti-Ro antibodies,anti-La antibodies,anti-Sm
antibodiesanti-nRNP antibodiesanti-Scl-70 antibodies, anti-histone
antibodies, antibodies to nuclear pore complexes, anti-centromere
antibodies,andanti-sp100 antibodies.
Anti-dsDNA antibody.
Diagnosis of Lupus nephritis, viral disease like parvo B19 and BK
virus.
Previously measured using radioimmunoassay (particularly the Farr
assay)
Nowadays measured using ELISA.
Immunofluorescence assay
target antigenCrithidia luciliae, a flagellated protozoa containing a dsDNAcontaining small organelle called a kinetoplast
Extractable
Nuclear Antigens
Anti-ENA testing
Sumayya
Antigens
Main antigens used in the laboratory for
disease detection are:
Ro
La
Sm
RNP
Scl-70
Jo1
Ouchterlony double
immunodiffusion
Agar gel immunodiffusion or passive double
immunodiffusion
Immunological technique
Involved in Identification, Detection and Quantification
The Technique
Principal: Anti-ENA
testing is based on the
binding of the antibodies
from the patient to the
antigens coated on a
microplate
Results
Signal Recognition
Particle
Anti-Signal Recognition Particle Auto-antibodies
What is it?
Abundant
Cytosolic
Universally conserved ribonucleoprotein
They target and bind to specific proteins in the EPR
Anti-SRP auto-antibodies
Presence is associated with severe acquired necrotizing
myopathies
Characterized with elevated serum Creatine Kinase levels,
severe muscle weakness and poor response to traditional
immunoresponsive therapy
Diagnosis: 3-4 months
Detection Methods
Indirect Immunoflourescence test (Not Specific)
Needs to be confirmed by an immunodot assay or protein
immunoprecipitation (Not Quantitative)
Immunodot Assay
Anti-Neutrophil Cytoplasmic
Antibody
A group of auto-antibodies
Mainly, IgG type
They are agianst cytoplams of neutrophils and monocytes
Detected in a number of auto-immune disorders
Particularly associated with vaculitis
ANCA Antigens
c-ANCA
Protinase 3
P-ANCA
Myeloperoxidase
In addition p-ANCA has its antibodies against Bacterial
permeability increasing Factor (BPI), Cathepsin G,
Elastase, Lactoferrin and lysozyme.
Others: HMG1, HMG2, alpha enolase, catalase and actin
e.tc.
Immunoglobulins
Cryoglobulin
Flow cytometry
cytokine studies
MHC studies
Cryoglobulin
Types of Cryoglobulin
Type I
Monoclonal immunoglobulins
IgM isotype
Multiple myeloma
Waldenstrom's macroglobulinemia
Lymphoproliferative disorders
Types of Cryoglobulin
Type II
Mixture of polyclonal igG and monoclonal igm
Type III
Combination of polyclonal igG and polyclonal igM.
Hepatitis C
Small vessel vasculitis
Detection Methods
If the cryoglobulin test is positive :
Protein electrophoresis
Immunofixation electrophoresis (IFE)
MHC/HLA
HLA studies
HLA-B27 is present in approximately 90% - 95% of white
patients with ankylosing
HLA-DR1 and HLA-DR4 increase the risk of polyarticular
juvenile idiopathic arthritis (JIA) in many populations.
HLA-DR3 and HLA-DR2 are associated with lupus in
Caucasian populations
Detection of HLA
Elisa
Gel electrophoresis
Polymerase chair reaction (PCR)
Cytokine studies
Cell signaling molecules that aid cell to cell
communication in immune responses
Stimulate the movement of cells towards sites of
inflammation, infection and trauma.
Cytokines that influence inflammation include il-1, il-6 and
tnf-alpha.
Detection
o DNA microarray studies
Rheumatoid Arthritis
Systemic Lupus Erythematosus
Multiple Sclerosis
Psoriasis
Systemic Sclerosis
THANKS