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Rapid and differential diagnosis of highly pathogenic avian influenza virus (HPAIV) subtype H5 is essential for the effective prevention and control of outbreaks caused by this pathogen. In this study, we describe a one-step multiplex real-time reverse transcription polymerase chain reaction (mRRT-PCR), using H5-, N1-, and N8-specific primers and probes, for differential detection of two HPAIVs (H5N1 and H5N8) and other H5-subtype AIVs. Using the mRRT-PCR assay, we were able to detect H5N1, H5N8, and other H5-subtype AIVs in a one-tube reaction, with high specificity; furthermore, using an in silico PCR program, we confirmed that this assay can detect nearly all H5, N1, and N8 genes of AIVs currently available in the Influenza Sequence Database. The limit of detection of the assay was determined to be as low as 100 copies/reaction for each target gene, and was comparable to limits of detection of previously reported mRRT-PCR assays. Thus, the mRRT-PCR assay described here can serve as a rapid and reliable differential diagnostic tool for the monitoring and surveillance of H5N1, H5N8, and other H5-subtype AIVs in countries where these pathogens are problematic.

Trichinellosis is a parasitic zoonosis that is of importance to public health; human trichinellosis usually occurs when improperly cooked pork or wild animal meat is consumed. The purpose of this study was to determine the nationwide seroprevalence of Trichinella infection in wild boar and domestic pig populations in Korea. Using ELISA, we detected no seropositivity among the serum samples of 2350 domestic pigs collected in 2013, indicating that the domestic cycles of Trichinella spp. have disappeared from the domestic pig population in Korea. In contrast, approximately 13% of the 434 wild boars hunted in 2013 were seropositive. Furthermore, the seroprevalence of six of the seven provinces was between 6.7% and 18.3%, indicating that Trichinella infection occurred in the wild boar population throughout the country. The results of this study suggest that Trichinella circulates in the wild boar population and could be transmitted from infected wild boars to other wildlife, domestic pigs, and humans in Korea. Therefore, we recommend continued surveillance of Trichinella infection prevalence in wild animals and an appropriate strategy to prevent human infection in Korea.