Publication: Preparation of Polyclonal Antiserum to the Recombinant TiLV-S8 Protein and Its Application in the Detection of Naturally Tilapia Lake Virus (TiLV) Infected Tilapia
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Issued Date
2023-07-01
Resource Type
eISSN
29850290
Scopus ID
2-s2.0-105005572588
Journal Title
Science Essence Journal
Volume
39
Issue
2
Start Page
79
End Page
95
Rights Holder(s)
SCOPUS
Bibliographic Citation
Science Essence Journal Vol.39 No.2 (2023) , 79-95
Suggested Citation
Sopa C., Wangman P., Ponpukdee N., Chaivisuthangkura P., Longyant S. Preparation of Polyclonal Antiserum to the Recombinant TiLV-S8 Protein and Its Application in the Detection of Naturally Tilapia Lake Virus (TiLV) Infected Tilapia. Science Essence Journal Vol.39 No.2 (2023) , 79-95. 95. Retrieved from: https://hdl.handle.net/20.500.14740/21043
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Abstract
Tilapia lake virus (TiLV) is classified as a negative-sense, single-stranded RNA virus in the family Amnoonviridae. It is an enveloped virus with 10 genomic RNA segments, each coding for a protein. TiLV causes disease in tilapia, and outbreaks can lead to significant economic losses for the tilapia aquaculture industry. In this study, the gene encoding the segment 8 protein of TiLV was cloned into the expression vector pET15-b and then transformed into Escherichia coli strain BL21. After induction, the recombinant TiLV-S8 protein (rTiLV-S8), with a molecular mass of 20 kDa, was expressed, purified, and used to immunize mice. The mouse antiserum against rTiLV-S8 protein demonstrated specific immunoreactivity for the viral protein, approximately 19 kDa in TiLV-infected fish tissues, as determined by Western blotting. According to the results of the dot blotting assay, the antiserum was about 80 times less sensitive than one-step RT-PCR in detecting TiLV in homogenates of infected fish samples and showed no cross-reaction with uninfected fish tissues, other common fish viruses, or prevalent bacterial species found in aquatic animals. Furthermore, this polyclonal antiserum could be employed to identify TiLV-infected fish in the field using dot blotting assay, and the results can be confirmed by immunohistochemistry.