DR.

Miswar Fattah, MSi

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

1. Adequate number 1. Something to be

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

technical expertise

time and money

 Novelty Ethical

1. Conrms or 1. Respect for

refutes previous persons (Informed
ndings consent)
2. Extends previous 2. Benecence
ndings (Risk/Benet
3. Provides new Analysis)
ndings 3. Justice (Selection
of subjects).
 an No
at v
ek k e
nd isti l it
Pe stat a s
BIOMARKER

Feasibility
Study
Cost

ur
St u at
D d i t er y
e sa Li ud
i n St
ClinVar
BIOMARKER

No
vel it
Feasibility a s
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

 an No
at v
ek k e
nd isti l it
Pe stat a s
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
 an No
at v
ek k e
nd isti l it
Pe stat a s
BIOMARKER

Feasibility
Study
Stu
De di
sa in
This pattern
only in
infection or
acute condition

Patter
concentration
of inammation
marker
different in
chronic
(Obesity,
Hypertension,
etc)
 an No
at v
ek k e
nd isti l it
Pe stat a s
BIOMARKER

Feasibility
Study
ur
St u at
D d i t er y
e sa Li ud
i n St
 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 certied 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

brinogen, platelets, and other circulating
proteins.

Cellular elements during the process of

coagulation can release inammatory
mediators that may affect cytokine levels
 an No
at v
ek k e
nd isti l it
Pe stat a s
BIOMARKER

Feasibility
Study
Cost

ur
St u at
D d i t er y
e sa Li ud
i n St
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-1
ADMA IL-2
Adrenalin Leptin
Aldosteron MCP-1
ANP MMP-2
Angiotensin II MMP-9
Asam Folat NO
BNP Noradrenalin
Endothelin PGF 1
E-Selectin PGI2
Fas Ligand PAPP-A
F2-Isoprostan PRA
ICAM-1 Pro-BNP
VCAM-1 sPLA2
Vitamin B12 TGF-1
TNF-
Tromboxan
Sitokin anti dan pro-inflamasi

IL-1 IL-10
IL-1 IL-17
IL-2 IFN
IL-4
IL-5
IL-6
IL-18
TNF
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
Angiopoietin-1
ICAM-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 1 HDL
ANP Small dense LDL (Direk)
Angiotensin II SAA
NO sPLA2
PRA LpPLA2
Obesitas

- Adiponektin Total - sHBG

- Adiponektin HMW, LMW - TGF-1
- Asam Lemak Bebas - Unconj. Estriol
- Collagen type IV - Visfatin
- Ghrelin total - Vaspin
- Leptin
- Leptin reseptor
- Obestatin
- Resistin
 Alzheimer
Stroke
amyloid
H-FABP
Isoprostan
IFN-
sPLA2
IL-1
IL-8 ApoE4
IL-10
MBP Saraf & Neurotransmitter
NSE Dopamin
IL-1
Protein S-100B
Serotonin
TGF-1 Endostatin
MMP-9 TNF-
BDNF
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
- 2 Glycoprotein 1
D-Dimer CRP Kuantitatif
DVV ICAM-1
Faktor VII
IFN-
Faktor VIII
MGP IL-1
PAI-1 IL-10
PAP-complex LBP
Protein C Aktif
TAFI Procalcitonin (PCT)
t-PA Protein C
Thrombomodulin TNF-
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 oor
Jl. Kramat Raya no. 150
Jakarta pusat
Telp : (021) 3144182 ext 712
Email : research.support@prodia.co.id
www.prodia.co.id