PENGEMBANGAN DETEKSI

MOLEKULER VIRUS
Sismindari

Faculty of Pharmacy UGM

6 April 2021
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 DETEKSI MOLEKULER VIRUS

Identifikasi molekul spesifik yang ada di

dalam Virus

Protein Asam Nukleat

Antibody Antigen RNA DNA

LFIA RT-Q-PCR Q-PCR

(Reverse (Quantitative
(Lateral Flow Immunoassay) transcriptase/Quantitative Polymerase Chain
PCR) Reaction)
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 DETEKSI MOLEKULER VIRUS

Identifikasi molekul spesifik yang ada di

dalam Virus

Protein Asam Nukleat

Antibody Antigen RNA DNA

LFIA RT-Q-PCR Q-PCR

(Reverse (Quantitative
(Lateral Flow Immunoassay) transcriptase/Quantitative Polymerase Chain
PCR) Reaction)
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 Tes PCR

Tes Antigen

Tes Antibodi

,, , , , , ,

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 -

COVID –19 antibody

specific against virus
antigen

Bentham Open
Oral COVID-19 Disclosing Test: A Novel Rapid Technique in Infection Diagnosis
Images may be subject to copyright.
https://benthamopen.com/FULLTEXT/TOPHJ-13-455/FIGURE/F2/

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 Monoklonal ANTIBODI
specific Nucleocapsid

ANTIGEN Membrane
(Nucleocapsid protein) glyco protein

Envelope
protein
Spike protein

SARS-CoV-2: An overview of virus structure,transmission and detection

https://diagnostics.roche.com/global/en/products/params/sars-cov-2-rapid-antibody-test.html

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1.How the SARS-CoV-2 EUA Antigen Tests Work
American Society for Microbiology, Aughttps://asm.org/Articles/2020/August/How-the-SARS-CoV-2-EUA-Antigen-Tests-Work. 31, 2020

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 CoKontrol

CoNegatif

Positif lemah

Positif

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 - à
Nama Urutan (5’ -3’) Panjang Amplikon
Gena RdRp/ nCoV_IP2
nCoV_IP2-12669Fw ATGAGCTTAGTCCTGTTG 17
nCoV_IP2-12759Rv CTCCCTTTGTTGTGTTGT 18 108 pb
nCoV_IP2-12696bProbe(+) AGATGTCTTGTGCTGCCGGTA [5']Hex 21
[3']BHQ-1
Gena RdRp/ nCoV_IP4
nCoV_IP4-14059Fw GGTAACTGGTATGATTTCG 20
nCoV_IP4-14146Rv CTGGTCAAGGTTAATATAGG 19 107 pb
nCoV_IP4-14084Probe(+) TCATACAAACCACGCCAGG [5']Fam [3']BHQ-1 19

Gena E / E_Sarbeco
E_Sarbeco_F1 ACAGGTACGTTAATAGTTAATAGCGT 18
E_Sarbeco_R2 ATATTGCAGCAGTACGCACACA 20 125 pb
E_Sarbeco_P1 ACACTAGCCATCCTTACTGCGCTTCG [5']Fam 20
https://www.who.int/docs/default-source/coronaviruse/real-time-rt-pcr-assays-for-the-detection-of-sars-
[3']BHQ-1
cov-2-institut-pasteur-paris.pdf?sfvrsn=3662fcb6_2
Posisi primer pada genome: nCoV_IP2/12621-12727, nCoV_IP4/14010-14116, Egene/26141-262530
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 Membrane glyco protein
Nucleocapsid protein

Envelope protein

Spike protein Primer target

SARS-CoV-2: An overview of virus structure,transmission and detection

https://diagnostics.roche.com/global/en/products/params/sars-cov-2-rapid-antibody-test.html

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 Specific Detection Systems
Taqman probes are “hydrolysis” probes

Polymerization
TaqMan®
Forward
R Probe Q
Primer
5’
3’ 5’

5’ 3’
5’
Reverse
R = Reporter dye Primer
Q = Quencher

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 Displacement

R
Forward
Q
Primer
5’
3’ 5’

5’ 3’
5’
Reverse
Primer

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 Hydrolysis

Q
5’
3’ 5’
5’ 3’
5’

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 Polymerization Completed

R Q

5’
3’ 5’

5’ 3’

5’

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 RNA

q PCR
cDNA

Analytical and Bioanalytical Chemistry (2021) 413:2311–2330.,

https://doi.org/10.1007/s00216-020-03046-0

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 RAPID ANTIBODI RAPID ANTIGEN PCR

Prinsip kerja Deteksi Antibodi Deteksi Protein Deteksi genom

(IgM & IgG) spesifik dari virus (RNA) virus
Sampel Darah Swab Nasopharynx Swab Nasopharynx

Waktu Analisis 15 menit 15 menit 3-5 jam

Sensitivitas Rendah Rendah Tinggi

Waktu sampling 1-3 minngu setelah Awal infeksi Awal infeksi

infeksi
Tingkat kesulitan rendah rendah tinggi

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 a si h
i ma k
Te r

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