Malaysian Journal of Analytical Sciences Vol 24 No 2 (2020): 179 - 187

 

 

 

 

DEVELOPMENT OF AN ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) FOR DOMOIC ACID DETECTION IN SELECTED SHELLFISH SEAFOOD USING α-AMINO-3-HYDROXY-5-METHYL-4-ISOXAZOLEPROPIONIC ACID (AMPA) RECEPTOR

 

(Pembangunan Imunojerapan Berpaut Enzim Berpesaingan Tak Langsung (ELISA) bagi Pengesanan Domoik Asid dalam Makanan Laut Bercengkerang Terpilih menggunakan Reseptor α-Amino-3-Hidroksi-5-Metil-4-Isoksazolepropionik Asid (AMPA))

 

Hafiza Mohamed Zuki*, Lai Yun Ru, Noor Sheryna Jusoh, Azrilawani Ahmad, Marinah Mohd Ariffin

 

Faculty of Science and Marine Environment,

Universiti Malaysia Terengganu,

21030 Kuala Nerus, Terengganu, Malaysia

 

*Corresponding author:  hafiza@umt.edu.my

 

 

Received: 20 November 2019; Accepted: 12 March 2020

 

 

Abstract

In this study, a selective and sensitive enzyme-linked immunosorbent assay (ELISA) method using ionotropic glutamate receptor of amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) was successfully developed in order to determine the presence of domoic acid (DA) in shellfish seafood. The optimum concentration of AMPA receptor and goat anti-mouse IgG alkaline phosphate (AP) were obtained at 1.3 ppm and 1.7 ppm, respectively. Absorbance response of the optimised assay towards DA recorded limit of detection (LOD) and limit of quantification (LOQ) at 0.72 ppm and 1.2 ppm, respectively. Cross-reaction with glutamic acid (GA) showed that the assay was less sensitive than DA, with cross-reactivity values of 12.96%. The competency of the immunoassay towards real samples analysis was studied using mussel and cockle. Detection of non-spiked samples recorded the presence of DA at 0.96 ppm for mussel and 0.56 ppm for cockle. Meanwhile, recovery of the spike samples with 3 ppm DA was found at 2.69 ppm for mussel and 2.60 ppm for cockle, with recovery percentages of 86.7% and 89.7%, respectively.

 

Keywords:  enzyme-linked immunosorbent assay, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, domoic acid, shellfish seafood

 

Abstrak

Dalam kajian ini, satu kaedah imunojerapan berpaut enzim berpesaingan tak langsung (ELISA) yang selektif dan sensitif dengan menggunakan reseptor glutamat ionotropik asid α-amino-3-hidroksi-5-metil-4-isoksazolepropionik (AMPA) telah berjaya dibangunkan untuk menentukan kehadiran asid domoik (DA) dalam makanan laut bercengkerang. Kepekatan optimum reseptor AMPA dan anti-tikus kambing IgG alkali fosfat (AP) masing-masing adalah didapati pada 1.3 ppm dan 1.7 ppm. Tindak balas penyerapan terhadap ujian optimum DA mencatatkan had pengesanan (LOD) dan had kuantifikasi (LOQ) masing-masing diperolehi sebanyak 0.72 ppm dan 1.2 ppm. Tindak balas silang dengan asid glutamat (GA) menunjukkan bahawa ujian GA adalah kurang sensitif daripada DA, dengan nilai tindak balas-silang 12.96%. Kecekapan imunoasai bagi analisis sampel sebenar telah dikaji menggunakan kupang dan kerang. Pengesanan sampel yang tidak dipaku mencatatkan kehadiran DA pada 0.96 ppm untuk kupang dan 0.56 ppm untuk kerang. Sementara itu, perolehan semula sampel yang dipaku dengan 3 ppm DA diperolehi pada 2.69 ppm bagi kupang dan 2.6 ppm bagi kerang, dengan peratusan pemulihan masing-masing sebanyak 86.7% dan 89.7%.

 

Kata kunci:  ujian imunojerapan berpaut enzim berpesaingan tak langsung, asid α-mino-3-hidroksi-5-metil-4-isoksazolepropionik, asid domoik, makanan laut bercengkerang

 

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