Osong Public Health and Research Perspectives

Original Article

Cloning and Expression of Recombinant Tick-Borne Encephalitis Virus-like Particles in Pichia pastoris: Seok-Min Yun, Young Eui Jeong, Eunbyeol Wang, Ye-Ji Lee, Myung Guk Han, Chan Park, Won-Ja Lee, WooYoung Choi; Osong Public Health Res Perspect. 2014;5(5):274-278. Published online October 31, 2014; DOI: https://doi.org/10.1016/j.phrp.2014.08.005

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Objectives
The purpose of this study was to verify the feasibility of using the glyceraldehyde-3-phosphate dehydrogenase (GAP) promotor based Pichia pastoris expression system to produce tick-borne encephalitis virus (TBEV) virus-like particles (VLPs).
Methods
The complementary DNA encoding the TBEV prM signal peptide, prM, and E proteins of TBEV Korean strain (KrM 93) was cloned into the plasmid vector pGAPZɑA, then integrated into the genome of P. pastoris, under the control of the GAP promoter. Expression of TBEV VLPs was determined by Western blotting using monoclonal antibody against TBEV envelope (E) protein.
Results
Recombinant TBEV VLPs consisting of prM and E protein were successfully expressed using the GAP promoter-based P. pastoris expression system. The results of Western blotting showed that the recombinant proteins were secreted into the culture supernatant from the P. pastoris and glycosylated.
Conclusion
This study suggests that recombinant TBEV VLPs from P. pastoris offer a promising approach to the production of VLPs for use as vaccines and diagnostic antigens.

Citations

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Review Article

Travel-Associated Chikungunya Cases in South Korea during 2009–2010: Go Woon Cha, Jung Eun Cho, Eun Ju Lee, Young Ran Ju, Myung Guk Han, Chan Park, Young Eui Jeong; Osong Public Health Res Perspect. 2013;4(3):170-175. Published online June 30, 2013; DOI: https://doi.org/10.1016/j.phrp.2013.04.008

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Abstract PDF

Objectives
Chikungunya (CHIK) has been classified as a communicable disease group IV in South Korea since late 2010. Based on this, we investigated the extent of imported cases of CHIK in dengue-suspected individuals returning from dengue-endemic regions.
Methods
A total of 486 dengue-suspected serum samples were screened for CHIK by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) analysis. Further RT-PCR-positive samples were used for the viral culture, and CHIK was subsequently confirmed by sequence analysis of the culture samples.
Results
Five out of 107 dengue-positive samples were found to be positive for CHIK and 15 out of 379 dengue-negative samples were found to be positive for CHIK by immunoglobulin M ELISA. Further, a CHIK virus was isolated from one of the two RT-PCR-positive sera by cell culture and confirmed by sequence analysis.
Conclusion
The present study documents the first evidence of travel-associated CHIK infection in South Korea. Considering the intense international traffic between countries, our finding emphasizes the urgent need for active patient and vector surveillance for timely response to reduce the introduction of CHIK in Korea.

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Article

Prevalence of Tick-Borne Encephalitis Virus in Ixodid Ticks Collected from the Republic of Korea During 2011–2012: Seok-Min Yun, Bong Gu Song, WooYoung Choi, Won Il Park, Sung Yun Kim, Jong Yul Roh, Jungsang Ryou, Young Ran Ju, Chan Park, E-Hyun Shin; Osong Public Health Res Perspect. 2012;3(4):213-221. Published online December 31, 2012; DOI: https://doi.org/10.1016/j.phrp.2012.10.004

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Abstract PDF

Objectives
In this study, we demonstrated that TBEV-infected ticks have been distributed in the ROK, combined with our previous results. These results suggest that TBEV may exist in the ROK, and H. longicornis, H. flava, and I. nipponensis may be potential vectors of TBEV. In addition, these results emphasize the need for further epidemiological research of TBEV.
Methods
We examined for the presence of RNA of TBEV by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) using ixodid ticks captured in 25 localities of 10 provinces. Ticks were collected by the flagging and dragging method or using sentinel BG traps at forests, grass thickets, and grassland. A total of 13,053 ticks belonging to two genera and four species were collected and pooled (1292 pools), according to collection site, species of tick, and developmental stage.
Results
Among 1292 pools, the envelope (E) protein gene of TBEV was detected using RT-nested PCR in 10 pools (3 pools of the 1,331 adult ticks and 7 pools of the 11,169 nymph ticks) collected from Gangwon-do province, Jeonrabuk-do province, and Jeju Island. The minimum infection rates for TBEV of Haemaphysalis longicornis, Haemaphysalis flava, and Ixodes nipponensis were 0.06%, 0.17%, and 2.38%, respectively. Phylogenetic analysis based on the partial E protein gene was performed to identify relationships between the TBEV strains. This showed that 10 Korean strains clustered with the Western subtype.
Conclusion
In this study, we investigated the prevalence of tick-borne encephalitis virus (TBEV) in ixodid ticks from various regions of the Republic of Korea (ROK) during 2011–2012 to identify whether TBEV is circulating and to determine the endemic regions of TBEV.

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