• Pediatric Infection and Vaccine
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• 제25권3호
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• pp.132-140
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• 2018

목적: 뎅기바이러스 감염 환자 발생은 해외 여행이 증가함에 따라 증가하고 있다. 최근 감염 경험이 없는 사람에게 백신을 접종하면 이후 야생 뎅기바이러스 감염 시 중증 뎅기열 증상을 보일 수 있다고 연구된 바 있다. 따라서, 본 연구는 한국의 다양한 연령군에서 뎅기바이러스에 대한 혈청역학을 연구하였다. 방법: 건강한 영아 98명(2개월-1세), 청소년 152명(13-19세), 성인 90명(20-50세) 및 노인 106명(65세 이상)에서 수집한 혈청 총 446명을 대상으로 하였다. 각 연령군의 뎅기바이러스 immunoglobulin G (IgG) 항체 검사를 ELISA을 통해 측정하였다. 또한 뎅기바이러스 IgG 항체 검사에서 양성 또는 equivocal을 보이는 혈청에 한하여 일본뇌염 바이러스의 IgG 항체를 검사하였다. 결과: 총 446명 검체 중, 청소년군에서 1명(0.2%)만 뎅기바이러스 항체 검사에서 양성으로 나왔다. Equivocal은 14명(3.1%)으로, 청소년군 10명과 노인군 4명이 해당하였다. 뎅기바이러스 IgG 양성이 나온 1명에서 일본뇌염 바이러스 또한 IgG 양성으로 나왔다. 뎅기바이러스 IgG equivocal이 나온 14명에서는 일본뇌염 바이러스 IgG 양성이 6명, equivocal이 3명이었고 음성은 5명이었다. 결론: 한국인에서 뎅기바이러스에 대한 항체 보유율은 매우 낮았다. 본 연구는 향후 뎅기열 예방을 위한 보건 정책 수립에 중요한 자료가 될 수 있을 것이며 향후 지속적인 혈청면역 평가도 필요할 것이다.

• 대한미생물학회지
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• 제1권1호
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• pp.3-3
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• 1958

• Journal of Preventive Medicine and Public Health
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• 제20권1호
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• pp.137-146
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• 1987

The following facts have been identified as a result of epidemiological trend and characteristic of Japanese Encephalitis in Korea for the last 20 years. First: The Epidemiological period which was ten-year and three-year in the past has been disappeared following the start of immunization program at 1970. Second: The Incidence rate was much higher in the south and West areas than northeast area of Korea. City and Province with the highest incidence rate was Chungcheong Nam Province and Cholla Buk Province. Third: Regardless of scope of prevalence, the main season that 90 percent of total incidence occurrs in one month from mid-August through mid-September. Fourth: The number of case by age was that 80 percent of total patients is children aged $3{\sim}15$. Recently there is an increase in the number of patients who are elderly people. Fifth: The study on the ecological conditions of mosquito including wintering and effectiveness of immunization for Japanese Encephalitis and duration on antibody should be done. Sixth: There has been no case of Japanese Encephalitis for the last three years since 1984 mainly due to disinfecting to eradicate mosquitos, immunization for vulnerable group of people aged $3{\sim}15$, individual precaution not to be bitten by mosquito, improvement of environment sanitation. While there has been no case of Japanese Encephalitis during last three years, there is possibility that Japanese Encephalitis becomes prevalent again anytime since its virus has been isolated continuously from the natural reservoirs.

• 대한수의학회지
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• 제10권2호
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• pp.59-66
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• 1970

Studies were conducted on the survey of HI antibody abortion and stillbirth from J.E. virus in swine for 1968-1969 in Korea HI antibody against J.E. virus showed postive in June or July and reached 80~100 percent of postive in August or September and J.E. virus strains were isolated from the brain material of the aborted piglets.

• 대한바이러스학회지
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• 제26권2호
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• pp.191-204
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• 1996

The characterization of the 5 Korean isolates (K96P10, K94P05, K91P55, K87P39, and K82P01) of Japanese encephalitis virus (JEV) was compared with JE virus prototype Nakayama-NIH (NKY-NIH) using prM/M and envelope gene sequences of the JEV genome and phylogenetic analysis. The antigenic analysis of these viruses were done by the cross-hamagglutination inhibition (HI) test using polyclonal antibodies against Korean isolates and NKY-NIH. The sequence homology of the Korean isolates and NKY-NIH ranged between 87.4 % - 95.6 % at the nucleotide level and between 98.2 % - 97.2 % at the amino acid level over the E nucleotides compared. Alignment of E protein amino acid sequences revealed that residue positions E89, E129, E221, E244, E327, E366, E459, and E477 characterized the Korean strains. According to phylogenetic analysis bases on the E nucleotide, there are at least 2 genetic types of JEV existing in Korea and Korean strains were distinct from NKY-NIH. However, the cross HI test results of all the Korean isolates were serologically no different from NKY-NIH strain.

• Journal of Microbiology and Biotechnology
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• 제22권11호
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• pp.1580-1587
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• 2012

Japanese encephalitis virus (JEV) envelope (E) protein holds great promise for use in the development of a recombinant vaccine. Purified recombinant E (rE) protein may be useful for numerous clinical applications; however, there are limitations in using the Escherichia coli expression system for producing high-quality rE protein. Therefore, in this study, the yeast expression system was used to generate the rE protein. For protein production using the yeast system, the full-length JEV E gene was cloned into Pichia pastoris. SDS-PAGE and immunoblotting analysis demonstrated that the rE protein had a molecular mass of 58 kDa and was glycosylated. The predicted size of the mature unmodified E protein is 53 kDa, suggesting that post-translational modifications resulted in the higher molecular mass. The rE protein was purified to greater than 95% purity using combined ammonium sulfate precipitation and a SP-Sepharose Fast Flow column. This purified rE protein was evaluated for immunogenicity and protective efficacy in mice. The survival rates of mice immunized with the rE protein were significantly increased over that of Hyphantria cunea nuclear polyhedrosis virus E protein (HcE). Our results indicate that the rE protein expressed in the P. pastoris expression system holds great promise for use in the development of a subunit vaccine against JEV.

• 대한수의학회지
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• 제57권1호
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• pp.31-36
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• 2017

Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

• 미생물학회지
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• 제30권2호
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• pp.115-123
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• 1992

박쥐 총 453 수의 일본뇌염 바이러스에 대한 HI 항체가를 조사하였던 바 1 :10 이상 양성률은 453 수 중 335 수(74.0%) 로 높은 양성률을 보였고 성별 항체보유율은 수컷이 70.0%(237수 중 166 수) 이고 암컷이 78.2%(216수 중 169수) 로 암켓이 약간 높게 나타났다. 박쥐 종류별 일본뇌염 바이러스 (Japanese Encephalitis Virus, JEV) 에 대한 HI 항체 양성률을 조사한 바 관박쥐 (Rhinolophus ferrumeguinum) 는 75.0%(360 수 중 34 수) 양셩이고 안주애기박쥐(Verpertilio superrans) 는 87.5%(24 수 중 21 수) 가 향체가를 보였고 큰수염박쥐 (Myotis mystatinus) 도 수가 적어 비교가 어려우나 항체보유률은 83.3%(12 수 중 10 수)가 양성이었다. JEV(Nakayama strain)을 한국산 야생박쥐 뇌내에 감염시켜서 되염 바이러스가 박쥐 뇌내 세포에서 증식하는지 여부를 확인하였다. 박쥐 뇌세포에서 JEV 항원과 바이러스의 감염입자를 전자 현미경으로 확인하였다.

• KSBB Journal
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• 제13권3호
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• pp.231-237
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• 1998

세계보건기구 (WHO)가 백신 생산에 권장하고 있는 표준세포 주인 Vero 세포에 약독화 일본뇌염바이러스인 SA14-14-2 ( (PDK)를 연속 계대배양을 통해 적응(adaptation)시켜, tIter가 $10^7$pfu/mL을 넘는 SA-14-14-2(Vero)을 분리하였다 바이러스 배양 최적온도는 $35^{\circ}C$이며, T -flask에서 배양된 바이러스의 최고 tIter는 감염 후 4일째에 $4\times10^7$ pfu/mL로 관찰되었다. 또한 무혈청배지에서도 바이러스 증식이 활발하여 2% 혈청이 보충된 정우와 거의 비슷한 바이라스 tIter를 보였다. 바이러스 대량 배양을 위해 roller bottle culture와 미 립 담체 플 이용한 spinner flask culture 가능성에 대하여 고찰하였다 바이러스 감염을 위한 미립담체에서의 Vera cell monolayer는 초기 세포 농도 $4\times10$ cells/mL로 접종하여 50 rpm에서 7일간 배양하여 얻을 수 있었다. 바이러스의 roller bottle 배양이 spinner flask 배양보다 바이러스 tIter변에서 2배 내지 3배 높 았고, $10^7$pfu/mL을 넘는 배양 기간도 하루 죄었다. 하지만 두 배양 방법 모두 T -flask 배양에서와 같이 무혈청 배지를 사용 하여도 바이라스 증식이 활발했고, 최고조의 tIter를 보이는 배 양기간은 감염 후 2일째로써 T -flask 배양에서 보다 2일 빨랐다. Roller bottle culture의 경우, 감염 후 3일부터 17일까지 2 일 간격으로 배양액을 무혈청 EMEM으로 100% 교체하면서 매 양을 지속한 결과 3일부터 9일까지 $10^7$pfu/mL을 념는 tIter가 유지되는 것이 확인되어 바이러스의 multi-harvest가 가능한 것 로 고찰되었다. 상기의 결과는 생산성 면에서 매우 유리한 결 과로 제품의 생산 단가플 낮추고 작업 노력을 절감하는 기대 효 과가 클 것으로 예측된다.

• 대한미생물학회지
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• 제3권1호
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• pp.43-49
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• 1968

Japanese encephalitis(JE) shows its explosive epidemicity in the temperate zone of Asia but little is known on the overwintering mechanism. One of the hypotheses on the overwintering mechanism is that the virus overwinters in the hibernating animals. There has been no report on the proliferation of JE virus(JEV) or antibody formation in the snakes. The purpose of this experiment is to explore the mutual relationship between JEV and snake and to clarify whether JEV proliferates and induce antibody formation in snakes. Three species of non-poisonous common snakes were employed. Precipitation test was carried out after injecting calf serum and, HI and neutralization tests were done by injecting JEV into the snakes. The gamma globulin fraction of pre- and post-injection serum were compared by paper chromatography. According to the results, precipitating antibody reaction to calf serum could be observed only at $4^{\circ}C$. It was failed to demonstrate HI antibody formation but neutralizing antibody could be detected in one of the 9 snakes. Although antibody could not be detected in test-tube, tile result of paper chromatography shows the remarkable increase of gamma globulin fraction after the injection. Above results are strongly indicating the antibody formation in the snakes.