ISSN: 1859 - 2872
Optimization of the Nested RT-PCR Protocol and Genetic Characterization of Hepatitis E Virus in Porcine Liver Tissue Samples Collected in Hanoi in 2025
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Abstract
Objective: This study aimed to optimize the conditions and evaluate the sensitivity and specificity of the Nested RT-PCR technique for detecting HEV. In addition, the infection rate and molecular features of HEV were investigated in porcine liver tissue samples collected in Hanoi in 2025. Subject and method: An experimental study was conducted in a molecular biology laboratory, combined with a cross-sectional descriptive survey of 298 porcine hepatic samples collected in Hanoi. Result: The Nested RT-PCR assay was successfully optimized with an annealing temperature of 560C and an extension time of 50 seconds for the outer cycle, and 540C with 40 seconds for the inner cycle. Among 298 liver specimens from pigs analyzed, 24 (8.05%) were identified as HEV-positive. Of these positive samples, 11 were successfully subjected to sequencing, enabling further molecular characterization. Phylogenetic analysis revealed that all strains belonged to genotype HEV-3, of which 9/11 were subtype HEV-3a, 1/11 HEV-3e, and 1/11 HEV-3f-like. Conclusion: The study established an optimized Nested RT-PCR protocol for HEV detection in porcine liver specimens. All positive samples were classified as genotype HEV-3, consistent with strains circulating in the region.
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References
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