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Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe

Foot-and-mouth disease (FMD) is known for its highly contagious properties among cloven-hoofed animals resulting in significant morbidity rates. Incursions of this disease have caused significant losses in affected countries in Southeast Asia and Africa, even within EU countries which resulted in si...

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Main Authors: Nordin, Nor Aina, Soon, Samson, Senawi, Jamaliah B., Jinin, Zurin Azlin M., Arshad, Siti Suri, Abd Rahaman, Yasmin, Azri, Farah Asilah
Format: Article
Published: Springer 2024
Online Access:http://psasir.upm.edu.my/id/eprint/114993/
https://link.springer.com/article/10.1007/s12010-024-05093-0?error=cookies_not_supported&code=15a63ff3-46b3-4652-b1db-b09713e3dfb4
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spelling my.upm.eprints.1149932025-02-14T02:12:03Z http://psasir.upm.edu.my/id/eprint/114993/ Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe Nordin, Nor Aina Soon, Samson Senawi, Jamaliah B. Jinin, Zurin Azlin M. Arshad, Siti Suri Abd Rahaman, Yasmin Azri, Farah Asilah Foot-and-mouth disease (FMD) is known for its highly contagious properties among cloven-hoofed animals resulting in significant morbidity rates. Incursions of this disease have caused significant losses in affected countries in Southeast Asia and Africa, even within EU countries which resulted in significant financial losses. This study is aimed at addressing existing limitations by creating a diagnostic method using aptamer-based assay. Three DNA aptamers were engineered to target the VP2 region of the FMD viral capsid protein. Since VP2 demonstrates a highly conserved amino acid sequence across serotypes, the specifically designed aptamers can detect different serotypes of the virus. Aptamers were evaluated against VP2 capsid protein, which was synthesized based on sequences from serotypes A, O, and Asia 1 of the FMD virus. After the recombinant VP2 capsid protein was developed, expressed, and refined, it was applied using enzyme-linked aptamer sorbent assay (ELASA) to determine aptamers’ binding capability. A similar test was further conducted with purified FMD virus from serotype A and serotype O. The ELASA results displayed a notable sensitivity in identifying the FMDV. Under optimized conditions, the aptamers have LOD as low as 0.11 ng/mL with LOQ as low as 0.34 ng/mL. The binding strength analyzed using the equilibrium dissociation constant (Kd) showed strong binding affinity at 3.092 ± 0.05 nM. Based on these findings, the method shows significant potential with high sensitivity and specificity for FMD virus detection assay. Springer 2024-11 Article PeerReviewed Nordin, Nor Aina and Soon, Samson and Senawi, Jamaliah B. and Jinin, Zurin Azlin M. and Arshad, Siti Suri and Abd Rahaman, Yasmin and Azri, Farah Asilah (2024) Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe. Applied Biochemistry and Biotechnology. ISSN 0273-2289; eISSN: 1559-0291 https://link.springer.com/article/10.1007/s12010-024-05093-0?error=cookies_not_supported&code=15a63ff3-46b3-4652-b1db-b09713e3dfb4 10.1007/s12010-024-05093-0
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
description Foot-and-mouth disease (FMD) is known for its highly contagious properties among cloven-hoofed animals resulting in significant morbidity rates. Incursions of this disease have caused significant losses in affected countries in Southeast Asia and Africa, even within EU countries which resulted in significant financial losses. This study is aimed at addressing existing limitations by creating a diagnostic method using aptamer-based assay. Three DNA aptamers were engineered to target the VP2 region of the FMD viral capsid protein. Since VP2 demonstrates a highly conserved amino acid sequence across serotypes, the specifically designed aptamers can detect different serotypes of the virus. Aptamers were evaluated against VP2 capsid protein, which was synthesized based on sequences from serotypes A, O, and Asia 1 of the FMD virus. After the recombinant VP2 capsid protein was developed, expressed, and refined, it was applied using enzyme-linked aptamer sorbent assay (ELASA) to determine aptamers’ binding capability. A similar test was further conducted with purified FMD virus from serotype A and serotype O. The ELASA results displayed a notable sensitivity in identifying the FMDV. Under optimized conditions, the aptamers have LOD as low as 0.11 ng/mL with LOQ as low as 0.34 ng/mL. The binding strength analyzed using the equilibrium dissociation constant (Kd) showed strong binding affinity at 3.092 ± 0.05 nM. Based on these findings, the method shows significant potential with high sensitivity and specificity for FMD virus detection assay.
format Article
author Nordin, Nor Aina
Soon, Samson
Senawi, Jamaliah B.
Jinin, Zurin Azlin M.
Arshad, Siti Suri
Abd Rahaman, Yasmin
Azri, Farah Asilah
spellingShingle Nordin, Nor Aina
Soon, Samson
Senawi, Jamaliah B.
Jinin, Zurin Azlin M.
Arshad, Siti Suri
Abd Rahaman, Yasmin
Azri, Farah Asilah
Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe
author_facet Nordin, Nor Aina
Soon, Samson
Senawi, Jamaliah B.
Jinin, Zurin Azlin M.
Arshad, Siti Suri
Abd Rahaman, Yasmin
Azri, Farah Asilah
author_sort Nordin, Nor Aina
title Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe
title_short Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe
title_full Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe
title_fullStr Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe
title_full_unstemmed Aptamer-based detection of foot-and-mouth disease virus using single-stranded DNA probe
title_sort aptamer-based detection of foot-and-mouth disease virus using single-stranded dna probe
publisher Springer
publishDate 2024
url http://psasir.upm.edu.my/id/eprint/114993/
https://link.springer.com/article/10.1007/s12010-024-05093-0?error=cookies_not_supported&code=15a63ff3-46b3-4652-b1db-b09713e3dfb4
_version_ 1825162410910023680
score 13.244413

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