DR.

Miswar Fattah, MSi

Research & Esoteric Laboratory Head
Prodia Clinical Laboratory
miswar.fattah@prodia.co.id
 “biomarker” is defined as a laboratory
measurement that reflects the activity of a
disease process
 Biomarkers : measurable characteristics that
reflect physiological, pharmacological, or
disease processes in animal or human
 Feasibility  Interesting

1. Adequate number of 1. Something to be

subjects passionate about
2. Adequate technical 2. Something to be an
expertise/markers expert in
3. Affordable in time 3. Something that
and money makes a difference
4. Manageable in scope in the world
Feasibility
 Feasibility

technical expertise

time and money

 Novelty  Ethical

1. Confirms or refutes 1. Respect for persons

previous findings (Informed consent)
2. Extends previous 2. Beneficence
findings (Risk/Benefit
3. Provides new Analysis)
findings 3. Justice (Selection of
subjects).
 Cost

BIOMARKER

Feasibility
Study
ClinVar
BIOMARKER

Feasibility
Study
 Analyte
 Sample type
 Sensitivity
 Availability : depend of our facility
 Single or multiplex
 Range of measurement
 Variation of results
 Quantitative or Qualitative
 Soluble receptors, soluble adhesion molecule,
Cytokine, interferon, chemokines : ELISA, RIA, luminex,
immunoturbidimetry, protein micro array
 Receptors, adhesion molecule, Cluster Differentiation
(CD) on cell or protein intracellular : Flowcytometry,
IHC
 Copy number of RNA on cell : Nucleic acid testing
 Serum, plasma, salive : ELISA, RIA, luminex,
immunoturbidimetry, protein micro array, nucleic acid
testing

 Direct Tissue : IHC, nucleic acid testing

 Cell : Flowcytometry, IHC, nucleic acid testing

BIOMARKER

Feasibility
Study
Konsentratsi biomarker

R positif R negatif

Berat penyakit/waktu
 Figure 1
Gitte Roende
(2014) 75, 551-558. doi:10.1038/pr.2013.252
 Scatter plot of peak mRNA values vs. homeostatic GC (o) amplitude for 2,092 of 100,000
parameter sets that have homeostatic mean mRNA values close to those produced by the
default parameters in Table 1.

Jeremy D. Scheff et al. Physiol. Genomics 2012;44:121-129

©2012 by American Physiological Society

 Quantitative : ELISA, Flowcytometry, nucleic
acid testing

 Qualitative
or semiquantitative : IHC,
Immunochromatography, nucleic acid testing
 Variation within run CV 20%
P=0,10
 Variation between run
 Variation between lot
 Biological variation
CV 10%
 Difference of case vs P=0,05
control must higher than
analytical variation
CV 5%
P=0,01

35 pg/mL 40 pg/mL
BIOMARKER

Feasibility
Study
This pattern
only in
infection or
acute condition

Patter
concentration
of inflammation
marker
different in
chronic
(Obesity,
Hypertension,
etc)
BIOMARKER

Feasibility
Study
 Research use Only (RUO)
 Laboratory Development Test (LDT)
 In vitro Diagnostic (IVD)

Approved Reagent by :
 CE mark (Europe)
 FDA Approved (USA)
 Reagent in research (RUO) different with routine
InVitro Diagnostic test.

 RUO test not internationally standardized

 IVD RUO

CRP, hsCRP IL-1,2,3,4, 6, 10 ,12, etc

Calprotectin TNF alpha

MPO NFKB

Fibrinogen sVCAM, sICAM, sP Selectine

Procalsitonin MCP-1

C3 & C4 IFN Gamma

 Reagent A

Reagent B

Reagent C

Need standardization before In vitro diagnostic test

Note : Using same reagent as a comparative literature
 Not all reagent have same measurement range

 Some biomarkers not to sensitive to detect low amount

of analyte

 Some biomarkers is bound with

protein/antibodi/soluble receptors

 Some biomarkers not active

 Cytokines from innate immune cells (MMP,
MPO etc) not be using serum, since there is a
possibility these get released during clotting

 Serum and plasma are not equivalen

 Plasma is preferable, generally collected in

EDTA, or heparin if it (and collection tubes) are
certified pyrogen-free.
 These cytokines are usually released in small
amounts, have very short plasma half lives, and
are restricted to the tissues.

 Serum preparation involves the removal of

fibrinogen, platelets, and other circulating
proteins.

 Cellular
elements during the process of
coagulation can release inflammatory
mediators that may affect cytokine levels
 Cost

BIOMARKER

Feasibility
Study
Low grade inflammation High grade inflammation

Moderate inflammation

High Sensitive

Non High Sensitive

High Grade Inflammation

Low Grade Inflammation

 ELISA
 RIA
 xMAP / Luminex
 Flowcytometry
 Immuno histochemistry
 Immuno turbidimetry
 Immuno chromatography
 Immuno chemiluminescence
 Nucleic Acid Testing
 Micro array
 Circulating markers
• Circulating DNA, mRNA & MicroRNA

 Multiplex Testing
• Multiple cytokine, chemokine, etc

 Applied new marker / reference interval

cut off for diagnosis
Pulmonologi (Blood or BAL)
– Progranulin (GP88) : Tissue & Circulating for non-
small cell lung carcinoma
– Collagen : PIIINP, CITP, MMP-9, & TIMP-1
– TGF
– CRP
– Fibrinogen
– CTC (Circulating tumor cell)*
– ADA
– YKL-40 (asma)
– cCK-18*
– Clara cell protein (CC16)
– SPLUNC1 (22-39)
– Surfactant protein A (SP-A)
– Surfactant Protein D (SP-D)
Disfungsi Endotel, Hipertensi & Resiko PJK
– ACE – IL-1b
– ADMA – IL-2
– Adrenalin – Leptin
– Aldosteron – MCP-1
– ANP – MMP-2
– Angiotensin II – MMP-9
– Asam Folat – NO
– BNP – Noradrenalin
– Endothelin – PGF 1a
– E-Selectin – PGI2
– Fas Ligand – PAPP-A
– F2-Isoprostan – PRA
– ICAM-1 – Pro-BNP
– VCAM-1 – sPLA2
– Vitamin B12 – TGF-b1
– TNF-a
– Tromboxan
Sitokin anti dan pro-inflamasi

– IL-1b – IL-10
– IL-1a – IL-17
– IL-2 – IFNg
– IL-4
– IL-5
– IL-6
– IL-18
– TNFa
– MCP-1
Plaque destabilization Oxidative stress

– MMP-1 – Isoprostan
– MMP-2 – MDA
– MMP-9 – MDA modified LDL
– MPO – Small dense LDL
– ICAM-1 – Ox-LDL
– VCAM-1
– P-selectin Apoptosis & Nekrosis
– E-selectin – HSP 70
– Anti HSP 70
– Cell Death Detection
– Fas Ligand
Plaque rupture Acute Phase reactant

– sCD40Ligand – CRP
– PlGF – sPLA2
– PAPP-A – Serum Amyloid A
– IL-6 – Fibrinogen
Marker Infark/ Iskemia Angiogenesis
Jantung
– Endostatin
– CD40L
– VEGF
– CK-MB
– ICAM-1 – Angiopoietin-1
– Lp(a) – Angiopoietin-2
– MMP-9
– MPO
– Myoglobin Myocardial dysfunction
– sPLA2
– Troponin I – BNP
– Troponin T – Pro BNP
– LpPLA2
Hipertensi Metabolisme Lipoprotein

– ACE – Apo A II
– ADMA – CETP
– FABP4
– Adrenalin
– FABP5
– Noradrenalin
– LCAT
– Dopamin – LDL Direk
– Aldosteron – Pre b1 HDL
– ANP – Small dense LDL (Direk)
– Angiotensin II – SAA
– NO – sPLA2
– PRA – LpPLA2
Obesitas

- Adiponektin Total - sHBG

- Adiponektin HMW, LMW - TGF-b1
- Asam Lemak Bebas - Unconj. Estriol
- Collagen type IV - Visfatin
- Ghrelin total - Vaspin
- Leptin
- Leptin reseptor
- Obestatin
- Resistin
Stroke Alzheimer

– H-FABP - b amyloid
– IFN-g – Isoprostan
– IL-1b – sPLA2
– IL-8 – ApoE4
– IL-10
– MBP Saraf & Neurotransmitter
– NSE – Dopamin
– Protein S-100B – IL-1b
– TGF-b1 – Serotonin
– MMP-9 – Endostatin
– BDNF – TNF-a
Infeksi
 HpSA
 Cag A
 Anti Dengue IgA
 Endocab (endotoxin core antibodies)
 ADA (Adenosine Deaminase)
 IgG anti-Measles
 Rotavirus Antigen
 Chlamydia pneumoniae IgG & IgM
 Infeksi (…lanjutan)

 CMV DNA (congenital inf)

 Staphylococcus aureus dan MRSA Tes
 Multiple Drug Resistance TB (MDR-TB) (Rifampisin
& isoniazid)  sputum
 M tuberculosis complex DNA (kualitatif)  sputum
 TB Quantiferon  darah
 HIV RNA (Realtime PCR)
 Dengue RNA serotyping (DEN1, DEN2, DEN3, DEN4)
 Dengue RNA (kualitatif)
 Entamoeba histolytica IgG
 Clostridium difficile Toxin A & Toxin B
Nutrisi & Pertumbuhan

 Transferin  25 Vitamin D
 Transferin reseptor (sTfR)  1.25 Vitamin D
 Prealbumin  EPO (erithropeitin)
 Feritin  Lactoferrin
 IGF-I
 IGF-II
 Vitamin A, E, C
 Asam Folat
 Vitamin B12
Koagulasi & Perdarahan Sepsis
- b2 Glycoprotein 1
– D-Dimer – CRP Kuantitatif
– DVV – ICAM-1
– Faktor VII – IFN-g
– Faktor VIII
– MGP
– IL-1b
– PAI-1 – IL-10
– PAP-complex – LBP
– Protein C Aktif
– Procalcitonin (PCT)
– TAFI
– t-PA – Protein C
– Thrombomodulin – TNF-a
– TSP-1
– vWF
– Tissue Factor
Keberhasilan dan efisiensi riset
mengenai biomarker ditentukan
oleh komunikasi yang efektif
antara Klinisi, laboratorium dan
Biostatistik

THANK YOU
Bagian Penunjang Penelitian
Laboratorium Klinik Prodia
Prodia tower 3rd floor
Jl. Kramat Raya no. 150
Jakarta pusat
Telp : (021) 3144182 ext 712
Email : research.support@prodia.co.id
www.prodia.co.id