Education

Dr. Miswar Fattah, MSi 1997 : SMAK Depkes Makassar

Makassar, 6th June 1978 2002 : Chemistry - UNHAS
2006 : Master of Science in Clinical Chemistry,
Biomedicine- UNHAS
2012 : Doctor of Medicine - UNHAS

Current position

1. Specialty & Research Laboratory Manager, Prodia Clinical Laboratory 2018- Now
2. PATELKI : Vice President 2017-Now & Member of Collegium PATELKI 2015 - Now
3. IACC: Member scientific committe, Indonesian Association for Clinical Chemistry 2013- Now
4. President of ASEAN Association of Clinical Laboratory Scientist (AACLS) 2018-2020
5. Corresponding Member Scientific Committee Asia Pacific Federation for Clinical Chemistry (APFCB)
2010 – Now
LATERAL FLOW TEST VS. LABORATORY BASED TEST
ANTIBODY SARS COV-2

DPP PATELKI Webinar

Jakarta, 28th Juni 2020

Dr. Miswar Fattah, MSi

Vice Presdient II PATELKI
Specialty & Research Laboratory
Prodia Clinical Laboratory
miswarfattah@gmail.com
KEY EVENTS IN THE 2019-NCOV OUTBREAK

3th march 2020

1st Case report in
Indonesia

Modiefied from Seah I et al. 2020. Eye, pp. 1–3

Rabi, F.A., Al Zoubi, M.S., Kasasbeh, G.A., Salameh, D.M., Al-Nasser, A.D., 2020. Pathogens 9, 231.
NAMING VIRUS AND DISEASE

Gorbalenya AE et al. 2020. Nature Microbiology. 5(4):536–44

 ROLE OF SCIENTIST DURING PANDEMIC

Predict potencial
Raising the Mechanisms of
emerging Profiling Virus
alarm Disease
disease

Prevent Next Develop

Contact tracers Develop Vaccine
outbreak Diagnostic Test
RACING TO DEVELOP COVID-19 TESTS
Electron
HE tissue Viral Culture NGS
microscope

HE tissue Electron microscope

Spesific Protein RT PCR,

ELISA
Antibody Isolation LAMP

Spesific Antibody RT PCR, LAMP

Rapid Lateral
Chemilumino
scence
“If you have a sequence today,
Flow
Autoanalyzer you have a PCR tomorrow”
Rapid Lateral Flow Chemiluminoscence Autoanalyzer
SARS COV-2 ANTIBODY
MAIN STRUCTURE OF CORONAVIRUSES
https://microbenotes.com/structure-and-genome-of-sars-cov-2/
DIFFERENT COVID‐19 CLINICAL COURSES AND TRAJECTORIES OF
ADAPTIVE IMMUNE RESPONSE AND VIRAL SHEDDING

Matricardi PM, Negro RWD, Nisini R. Pediatric Allergy and Immunology. 02 May 2020
DIFFERENT TYPE OF ANALYTE LABORATORY TESTING RELATED COVID-19

RNA Host Respons

Antigen Antibody Potencial
ORFla/b Gene CBC
N Protein IgM CRP
Succebility
N Gene
E Gene S Protein IgG D Dimer ACE2 Gene
S Gene IgA SGOT HLA Gene
Albumin
LDH, etc

rRTPCR, LAMP, ELISA, Immuno- ELISA, Immuno- Enzymatic, Genotyping

chromatography, colorimetry, microarray, RTPCR,
NGS chromatography flowcytometry,
Chemiluminoscence Sanger Seq, NGS
immnoassay impedance
TESTS FOR SARS–COV-2/COVID-19 AND POTENTIAL USES
Report from the American Society for Microbiology COVID-19 International Summit, 23 March 2020: Value of
Diagnostic Testing for SARS–CoV-2/COVID-19

R. Patel et al., mBio. 11 (2020), doi:10.1128/mBio.00722-20.

IFCC Information Guide on COVID-19 - IFCC. www.ifcc.org
Ejazi SA, Ghosh S, Ali N. 2020
https://microbenotes.com/structure-and-genome-of-sars-cov-2/
https://microbenotes.com/structure-and-genome-of-sars-cov-2/
THE HUMAN ANTIBODY LATCHING ONTO THE RECEPTOR BINDING
DOMAIN (RBD) OF THE SPIKE PROTEIN OF SARS-COV-2

UNC School of Medicie

Iwasaki A, Yang Y. 2020. Nature Reviews Immunology. 20(6):339–41
Role of Antibody
telated SARS
COV-2
 PROTEIN
IMMUNODOMINANCE
OF SARS-COV-2-
SPECIFIC CD4+ AND
CD8+ T CELLS IN
COVID-19 CASES AND
UNEXPOSED DONORS

Grifoni et al., 2020, Cell 181,

1489–1501, June 25, 2020
ANTIBODY KINETICS

Ab N protein & Anti S2 Ab Anti S1 subunit

RBD earliest to timelag of 2 days are the last to be
be detected after N & RBD detected
(100%) 13 days
after symptom
onset
The role of serologic testing in COVID-19 Diagnosis, Mario Plebani, june 2020
FIVE DIFFERENT BETACORONAVIRUSES WITH POTENTIAL
FOR CROSS-REACTIVITY

Hicks J, Klumpp-Thomas C, Kalish H, Shunmugavel A, Mehalko J, et al. 2020. medRxiv.2020.06.22.20137695

SARS-COV-2 SIGNAL INTENSITY COMPARED WITH SIGNAL INTENSITY
OF OTHER BETACORONAVIRUSES IN PANDEMIC HOT-SPOT
COMMUNITY BLOOD DRAWS

Hicks J, Klumpp-Thomas C, Kalish H, Shunmugavel A, Mehalko J, et al. 2020. medRxiv.2020.06.22.20137695

ANTI-SARS-COV-2 ANTIBODY KINETICS AND SENSITIVITY
LEVELS OF IGM, IGA, AND IGG ANTIBODIES AGAINST SARS-COV-2
IN PLASMA SAMPLES AFTER SYMPTOM ONSET

L. Guo et al., Clin. Infect. Dis. (2020), doi:10.1093/cid/ciaa310.

L. Guo et al., Clin. Infect. Dis. (2020), doi:10.1093/cid/ciaa310.
ANTIBODY IGG & IGM IN SEVERE COVID-19 PATIENTS

J. Zhao et al., Clin Infect Dis, doi:10.1093/cid/ciaa344.

 WHY WE NEDD SEROLOGICAL TEST?

Improve Screen particular Chracterize efficacy

Define & Monitor
knowlegde sub Population & of containment
the extent of virus
immuneresponse Identify Disease mesures at local &
spread
to SARS COV-2 prevalence Global

Identify individual
Combine rRT-PCR
Screen Diagnosis for later infected but
for more accurate
convalescent sera stages of infection suffered only
diagnosis
minor symtoms

The role of serologic testing in COVID-19 Diagnosis, Mario Plebani, june 2020
UNKNOW RELATES SEROLOGY

Likelihood of
Kinetics Level & duration
protection agains
antibodies of immunity
reinfection
TEST TO DETECT SARS COV-2 ANTIBODY

Point of Care Laboratory Based Neutralizing Antibody

(POCT): Test: Detection:
Lateral flow devices that ELISA or CLIA detect IgG,
Determine the functional
detect IgG or IgG & igM or IgM and IgA separately or ability of antibodies to
Total Ab. Whole blood, combined as total Ab prefent infection of virus
serum, plasma, saliva in vitro. VNT: virus
neutralization test; pVNT
presudovirus
neutralization test
COVID-19 Testing Project. https://covidtestingproject.org
Lateral Flow SARS COV-2
Antibody
 LATERAL FLOW BASED TEST

LFT band intensity

Reader training is key Agreement between score showed direct
to realiable LFT IgG & IgM ELISA correlation with ELISA
performance 75,8 – 85,7% signal/cut-off (S/CO)
values

Whitman JD, Hiatt J, Mowery CT, Shy BR, Yu R, et al. 2020. medRxiv. 2020.04.25.20074856
THE FOUR ANTIBODY RAPID TESTS FOR DIAGNOSIS OF (COVID-19)

Wu J-L, Tseng W-P, Lin C-H, Lee T-F, Chung M-Y, et al. 2020. Journal of Infection. 0(0):Published:June 15, 2020
THE FOUR ANTIBODY RAPID TESTS FOR DIAGNOSIS OF (COVID-19)

Wu J-L, Tseng W-P, Lin C-H, Lee T-F, Chung M-Y, et al. 2020. Journal of Infection. 0(0):Published:June 15, 2020
AGREEMENT OF SEROLOGICAL ASSAYS FOR SARS-COV-2

Whitman JD, Hiatt J, Mowery CT, Shy BR, Yu R, et al. 2020. medRxiv. 2020.04.25.20074856
Krammer F, Simon V. 2020. Science. 368(6495):1060–61
IMMUNOASSAY TARGET
Abbott Architecs (CLIA) • Nucleocapsid phophoprotein (IgG)

Diasorin Liason (CLIA) • Recombinan S1/S2 Spike p (IgG)

Euroimmune (ELISA) • S1 domain of spike (IgA, IgG)

SNIBE maglumi (CLIA) • S1/S2 Spike p + N Protein (IgM, IgG)

Wantai (ELISA) • RBP domain Spike (total Ab IgM)

Roche Cobas (ECLIA) • Nucleocapsid (total Ab)

Siemens Advia (CLIA) • S1 RBP domain spike (total Ab)

Ortho (CLIA) • S1 spike total Ab & Spike (IgG)

Mindray (CLIA) • recombinant N-Protein & Spike (S) Protein (IgG & IgM)
The role of serologic testing in COVID-19 Diagnosis, Mario Plebani, june 2020
PERFORMANCE CHARACTERISTICS OF SEROLOGY KITS
VERSUS THE RESULT OF PCR

Herroelen PH, Martens GA, Smet DD, Swaerts K, Decavele A-S. 2020. medRxiv. 2020.06.09.20124719
SEROLOGICAL SIGNATURES OF SARS-COV-2 INFECTION:
IMPLICATIONS FOR ANTIBODY-BASED DIAGNOSTICS

Rosado J, Cockram C, Merkling SH, Demeret C, Meola A, et al. 2020. medRxiv. 2020.05.07.20093963
SENSITIVITY AND SPECIFICITY TARGETS FOR MULTIPLE
BIOMARKER

Rosado J, Cockram C, Merkling SH, Demeret C, Meola A, et al. 2020. medRxiv. 2020.05.07.20093963
KINETICS OF IGA (ELISA) AND IGM (CLIA) OF PATIENTS MONITORED
FROM THE ONSET OF SYMPTOMS (FEVER)

Padoan A, Sciacovelli L, Basso D, Negrini D, Zuin S, et al. 2020. Clinica Chimica Acta. 507:164–66
LFIA VS CLIA VS. ELISA

IgG-IgM based LFIA

IgG-IgM based CLIA

IgG-IgM based ELISA

A Systematic and Meta-Analysis Review on the Diagnostic Accuracy of Antibodies in the Serological Diagnosis of COVID-19. 2020.
J. Zhao et al., Clin Infect Dis, doi:10.1093/cid/ciaa344.
THE POSITIVE DETECTION RATE IS SIGNIFICANTLY INCREASED
WHEN COMBINED IGM ELISA ASSAY WITH PCR

L. Guo et al., Clin. Infect. Dis. (2020), doi:10.1093/cid/ciaa310.

A DETAILED PROTOCOL FOR A SEROLOGICAL ASSAY TO DETECT
SARS-COV-2 SEROCONVERSION IN HUMANS: ANTIGEN
PRODUCTION AND TEST SETUP

Stadlbauer D, Amanat F, Chromikova V, Jiang K, Strohmeier S, et al.

2020. Current Protocols in Microbiology. 57(1):e100
SARS COV-2 NEUTRALIZATION TEST

https://www.businesswire.com/news/home/20200413005181/en/%C2%A0Celltrion-Completes-Neutralisation-Test-Candidate-Monoclonal-Antibodies
RATIONAL USE OF SEROLOGICAL TESTING

Understand the Collect data whether &

Undestand the
presense of SARS COV2 How antibodies confer
prerformance of test
in population immunity (degree of
(Sensitivity & Specifity)
(Prevalence) persistance)
STRATEGY OF TESTING

Maxiize specificity & PPV

Choosing a test with greater than 95% specificity

Focus testing with high pre test probability

Orthogonal testing algorithm (positive result tested with a second test)

Verify Emergency Use Authorization (EUA) SARS-CoV-2 Antibody Tests. ASM.org. www.asm.org
EXAMPLES OF INTERPRETIVE “VALUE-ADDED” COMMENTS FOR
SARS-COV-2 SEROLOGIC ASSAYS

Verify Emergency Use Authorization (EUA) SARS-CoV-2 Antibody Tests. ASM.org. www.asm.org
POPULATION IMMUNITY
Preliminary results of
first seroepidemiological
population studies in
EU/EEA Member States
and the UK from public
sources, as of 8 June
2020

Immune responses and immunity to SARS-CoV-2. European Centre for Disease Prevention and Control. www.ecdc.europa.eu
 Viral RNA Sequence
Antigens profiling
Viral protein efficacy
safety
Toxicity
doses Pabrication
Stability
Clinical Trial

store

The race to produce a vaccine

Increase volume of Testing before Vaccine ready

Herd immunity
TERIMA KASIH