Gene Expression of Toll-Like Receptors (TLR)-2 and-3 in QPCR-Confirmed Blood Samples Infected with African Swine Fever Virus (ASFV)

Date

2024

Degree

Doctor of Veterinary Medicine

College

College of Veterinary Medicine (CVM)

Adviser/Committee Chair

Cherry P. Fernandez-Colorado

Co-adviser

Mark Joseph M. Desamero

Abstract

Background: African swine fever (ASF) is a viral hemorrhagic disease affecting both domestic and wild pigs which causes socio economic losses due to high mortality and morbidity rates. There is no commercially available vaccine yet in the market, but vaccine trials are actively being conducted in other countries. Understanding how the host's immune system responds to ASF infection is critical not only in controlling AS outbreaks but also in developing diagnostic test kits, targeted control measures and effective vaccines. Hence, this study was conducted to determine patterns of activation of immune-related genes, specifically toll-like receptors (TLR)-2 and -3 in whole blood samples of ASF-infected pigs using qRT-PCR. Methodology: ASFV-positive whole blood samples were obtained from local government units (LGUs) or provincial veterinary offices (PVOs) which collected the samples from ASFV-infected farms for the AS monitoring and surveillance program of their respective areas of jurisdiction. The blood samples were subjected to further qPCR confirmation prior to gene expression analysis. Confirmation of ASFV positive diagnosis was subsequently performed following the recommended procedures outlined in the WOAH Terrestrial Manual on African Swine Fever Diagnosis. Afterwards, gene expression levels of TLR-2 and -3 were quantified using qRT-PCR following the 2-△△ Ct method. Results: In this study, only blood sample total RNAs with good quality were subjected to qRT-PCR for the gene expression analysis. The mRNA expression levels of TLR-2 and TLR-3 were highly significant as compared to the corresponding mRNA expression level of the negative control (NC). TLR-2 highly increased by 4137.32-fold, while there was 5338.25-fold increase in TLR-3 mRNA expression level. Moreover, TLR-3 mRNA expression level was higher by 1200.93-fold as compared to TLR-2 expression level. Conclusion: Overall, the findings of this study showed the specific and highly significant upregulation of TLR-2 and -3 mRNA transcript levels in ASFV-infected blood samples indicating the active involvement of the TLR signaling pathway in the host response against the virus

Language

English

Location

UPLB College of Veterinary Medicine (CVM)

Notes

Awards: Best Undergraduate Thesis Award in Veterinary Immunology

Document Type

Thesis

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