| dc.description.abstract | Introduction: Hepatitis E virus (HEV) poses a significant public health concern, with
genotypes 1 and 2 causing acute and chronic hepatitis in humans, and genotypes 3 and 4
exhibiting zoonotic potential through undercooked meat consumption and animal
contact. This thesis aimed to optimize a PCR protocol for sensitive and specific
detection of HEV RNA in fecal samples from domesticated wild boars, a potential
reservoir for zoonotic HEV strains.
Method: RNA was extracted and cDNA was synthesized from 27 domesticated wild
boar fecal samples collected in southern Vietnam. An established protocol was
optimized for annealing temperature, primer concentration, and magnesium chloride.
The optimized protocol was then evaluated for specificity, sensitivity, and performance
on real fecal samples.
Result: Optimization involved increasing the annealing temperature to 57.4°C and
adjusting primer and MgCl₂ concentrations. Optimization yielded high specificity,
sensitive detection (approximately 11.8ng/µL), and a promising 48.2% detection rate in
wild boar samples (13 out of 27 samples), suggesting potential high HEV prevalence.
Conclusion: A protocol for HEV detection in wild boars was optimized, achieving high
specificity, sensitive detection, and a promising 48.2% detection rate in initial validation.
However, confirmation of these positive results using established nested PCR and more
accurate DNA quantification methods is necessary for definitive validation. This work
provides a valuable tool for future studies investigating HEV prevalence in domesticated
wild boars and their potential role in HEV epidemiology. | en_US |
