• Title/Summary/Keyword: neutralizing antibody test

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Serological evidence on the persistence of porcine epidemic diarrhea virus infection (돼지 유행성 설사병(porcine epidemic diarrhea)의 상재화에 대한 혈청학적 증명)

  • Park, Bong-kyun;Han, Kyung-soo;Lyoo, Kwang-soo;Kim, Jun-young;Jeong, Hyun-kyu
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.818-822
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    • 1998
  • The persistence of porcine epidemic diarrhea virus(PEDV) infection was demonstrated in 7 swine farms employing continuous pig flow management even after seasonal outbreaks. Clinically, sporadic postweaning diarrhea was a major concern in those farms. Subsequently circulatory antibody detection using serum neutralizing test made useful for confirmation of PEDV persistent infections. The persistence of PEDV in the premise might have induced recurrence over the period of time.

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Evaluation of stability and potency of live attenuated rinderpest vaccine of lapinized-avianized tissue culture strain origin for the establishment of expiration period (가토화·계태화 우역 생백신의 유효기간 설정을 위한 안정성 및 효능 평가)

  • Yeh, Jung-Yong;Kim, Hyun-Ju;Nah, Jin-Ju;Park, Jee-Yong;Lee, Ji-Hye;Seo, Hyun-Ji;Kweon, Chang-Hee;Cho, In-Soo;Moon, Jin-San
    • Korean Journal of Veterinary Research
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    • v.49 no.3
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    • pp.201-205
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    • 2009
  • In this study, we evaluated the stability and potency of live attenuated rinderpest vaccines of lapinized-avianized tissue culture strain origin, which had been produced annually from 2005 to 2008. When immune responses to the vaccines were evaluated using two Holstein calves weighing 100~150 kg, neutralizing antibody titer of 1 : 16 was induced at 21 days post vaccination. When calves were also inoculated with vaccines lots that had been stored for 39 months at ${4^{\circ}C}$, same level of antibody titer was observed. Using the virus titer test, we found that all batches of the vaccine that had been kept for 3, 10, 15, 22, 27, 34, 39, and 45 months showed no significant loss of titers, and fulfilled the requirement necessary ($\geq$ 3 $logTCID_50$) to be used as the national rinderpest vaccine reserve in Korea. In this study, we demonstrated that stability and potency of the rinderpest vaccines were maintained over three years when kept at ${4^{\circ}C}$ storage. This indicates that it maybe feasible to extend the expiration period of this vaccine from one year to three years.

Immunocytochemical identification of Aujeszky's disease viral antigens in nasal and circulating white blood cells from living pigs for early diagnosis (오제스키병의 생체 조기진단을 위한 돼지 비세포 및 순환백혈구내 면역세포화학적 바이러스 항원동정)

  • Moon, Oun-kyong;Cho, Hee-tack;Kim, Soon-bok;Sur, Jung-hyang
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.241-247
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    • 1993
  • Forty day old piglets were intranasally inoculated with 2ml of Aujeszky's disease virus (NYJ-1-87 strain, $10^{7.0}$ $TCID_{50/0.2}ml$), and the viral antigens were detected in nasal and circulating white blood cells for 20 days after inoculation by immunocytochemical method. Antibody titers in the blood were also detected by neutralizing test and Aujeszky's disease serodiagnostic kit(Choong Ang) in this periods. 1. Viral antigens were detected by the immunocytochemical technigue, and positive reactions were observated in nasal cells from the 2nd to the l0th days after inoculation and circulated white blood cells from the 4th to the 12th days after inoculation. 2. In neutralization test antibodies levels showed titers of 2 on the 8th day, 8 on the l5th day, 16 on the 18th day and 32 on the 20th day after inoculation. In serodiagnostic kit test positive reactions were observed after the 15th day after inoculation.

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Production of Monoclonal Antibody to Newcastle Disease Virus and its Diagnostic Use (뉴켓슬병 바이러스에 대한 단크론성 항체생산 및 진단에 이용)

  • Chung Ok Choi;Chung Gil Lee;Sung Man Cho;Jin Soo Na;Soo Hwan An;Joon Hun Kwon
    • Korean Journal of Poultry Science
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    • v.15 no.3
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    • pp.207-210
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    • 1988
  • A total of 3 hybridoma clones producting menoclonal antibody (MCA) against Newcastle disease virus(NDV) was raised by cell fusion method. The MCAs did not cross react against other avian or mammalian viruses tested. However, these antibodies reacted with all strains of velogenic and lentogenic NDVs tested indicating that they are unable to discriminate the possible antigenic differences among NDVs. All. the MCAs were classified as IgG type and did not show neutralizing and hemagglutination inhibition (HAI) activity except one clone which has low HAI activity. One of these MCA raised in mouse ascites revealed the titer of $10^6$ by indirect immunofluorescent antibody (IFA) test Using the MCA, virulent NDV could easily be detected from tracheal and conjunctival smears made 2 to 3 days after experimental infection.

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Discrimination of Hantaviruses from the Tissues of Infected Hamsters to 5 Different Serotype Hantaviruses by Nested RT-PCR using Hantavirus Serotype Specific Primers (한타바이러스 혈청형 특이 Primer를 이용한 Nested RT-PCR 방법으로 5가지 혈청형 한타바이러스에 감염된 햄스터 조직에서 바이러스 검출)

  • Chu, Yong-Kyu;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.49-57
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    • 1997
  • We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

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Induction of Neutralizing Antibodies by Recombinant Nucleocapsid Protein (N) of Hantaan Virus: Potentiality and Implications (한탄바이러스의 유전자 재조합 내피단백질에 의한 중화 항체의 유도)

  • Noh, Kap-Soo;Hong, Sun-Pyo;Shin, Young-Cheol;Lee, Sung-Hee;Kim, Hyun-Su;Choi, Cha-Yong;Yao, Zhi-Hui;Kim, Soo-Ok;Yoo, Wang-Don
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.205-214
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    • 1996
  • 한탄바이러스의 내피 단백질 (N)은 이 바이러스에 대한 중요한 항원으로 작용하지만 신증후출혈열 예방과 관련된 작용은 명확히 알려져 있지 않다. 본 연구는 이러한 내피 단백질이 한탄바이러스에 대한 중화 항체를 유도할 수 있는가 하는 관점에서 수행되었다. 한탄바이러스의 내피 단백질을 대장균에서 용해된 형태로 발현하고 이를 단클론 항체를 이용한 면역친화 컬럼으로 분리 정제하였다. 정제된 내피 단백질을 기니픽에 면역하여 항혈청을 얻고 이것의 한탄바이러스에 대한 중화능력을 중화항체 플락 감소법 (plaque reduction neutralization test)을 이용하여 조사한 결과 최고 1:160의 중화능이 있음을 관찰하였다. 이는 한탄바이러스의 내피 단백질이 중화 항체를 유도할 수 있는 epitopes을 가지고 있음을 의리하며 이러한 생각은 본 연구에서 수행한 면역침강법과 N 단백질에 대한 단클론항체를 이용한 면역친화법을 통한 한탄바이러스의 정제 실험 결과에서도 뒷받침되고 있다.

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Persistency of Neutralizing Antibody to Inactivated Mouse Brain Derived Nakayama Japanese Encephalitis Vaccine and Current Observations of Booster Vaccination and Adverse Events (일본뇌염 사백신 중화항체 지속률과 부작용에 대한 연구)

  • Sohn, Young Mo;Park, Ji Ho;Lee, Jin Soo;Roh, Hye Ok;Ki, Moran;Choi, Bo Yul;Kim, Young Ho
    • Pediatric Infection and Vaccine
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    • v.8 no.2
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    • pp.150-159
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    • 2001
  • Purpose : We need to reconsider booster vaccination schedule of Japanese encephalitis vaccination. To do that we evaluate the long-term immunogenicity and the incidence of adverse events with inactivated mouse brain derived Nakayama Japanese encephalitis vaccine. Methods : We tested neutalizing antibody for 311 elementary school students by plaque reduction neutralizing test(PRNT) at USAMC-AFRIMS(United States Armed Forces Research Institute of Medical Science/Department of Virology). We evaluated vaccine related adverse events by spontaneous reporting prospectively among 15,487 vaccinees who were vaccinated at public health center and 2,277 elementary school students who were immunized previously by a questionnaire and school health record. Results : According to the time interval from the last booster injection of 311 children, PRNT antibody titers gradually decreased as the interval increased; 239 mIU/mL, 188 mIU/mL, 134 mIU/mL, 49 mIU/mL each at 6, 18, 30, 42 months after the last booster injection. The seropositivity rates were 98%, 99%, 95.6%, 71.4% each at 6, 18, 30, 42 months after the last booster injection. There were 21(0.13%) cases with systemic reactions among 15,487 vaccinees who had visited the hospital by prospective passive reporting system at public health center. According to the questionnaires and school health records in elementary school students, local induration and pain were 17.4% and 14.8%, respectively. Systemic reactions including fever, vomiting, rash were reported in few cases. Conclusion : Biannual booster vaccination that has been recommended so far should not be necessary. Surveillance for adverse events with inactivated mouse brain derived Nakayama vaccine should be strengthened to better assess the number of cases and reactions associated with immunization.

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Studies on the Duration of Immunity and Production of Antibody following Immunization with Inactivated Killed Japanese Encephalitis Vaccine (일본뇌염 백신 접종후 항 일본뇌염 항체의 생성율과 지속적인 면역반응에 대한 연구)

  • Cho, H.W.;Nam, J.H.;Lee, H.D.;Koh, H.C.;Kim, J.J.;Kim, E.J.;Lee, Y.S.;Lu, J.J.
    • Pediatric Infection and Vaccine
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    • v.4 no.1
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    • pp.116-125
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    • 1997
  • Purpose : Studies on the duration of immune response against Japanese encephalitis virus from recipients with JE vaccine (Nakayama-NIH strain) in Korea. Methods : To determinate the immune response and the duration of antibody against JE vaccine, 213 students were examined since 1994 using hemmaglutination inhibition test and plaque reduction neutralization test (PRNT). Results : 24 months after the first vaccination, haemmaglutination inhibition and neutralizing antibody maintained from the recipients 63.4% (>1:20) and 100% (>1:20), respectively. In April 1996, one dose booster to the same recipients those who were vaccinated in 1994, the GMT antibody for HI and PRNT titer were both increased from 1:11.6 to 1:13.2 and 1:275.7 to 1:348.1, respectively, after 6 months booster (after 30 months from the initial vaccination). This results showed that the antibody from the active immunity could be maintained more than 12 months after the initial vaccination. On the basis of these results, inactivated killed JE vaccine (Nakayama-NIH strain) using for preventing against JE purpose seems to produce antibody enough to protect against JE at present. Conclusions : Along with the results of this study demonstrating duration of antibody, the active immunization could be maintained as long as by initial vaccination of 2 doses, a single dose of booster vaccination made during a period of 1 month to 12 months and the successive booster vaccination by 2 or 3 year intervals. However, the immunization schedule should be concerned with both epidemiology of disease and the immune response of vaccinated individuals.

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An Immunoserological Study of Vaccine Against Haemorrhagic Fever with Renal Syndrome (신증후출혈열 백신의 면역혈청학적 연구)

  • Woo, Young-Dae;Chu, Yong-Kyu;Baek, Luck-Ju;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.11-18
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    • 2000
  • Since $Hantavax^{TM}$, formalin inactivated Hantaan virus vaccine (10,240 ELISA units/ml), has been developed in 1990 to prevent against haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan or Seoul virus, it has been commercially available in Korea. Twenty-one healthy people were booster shot once and twice after primary basic vaccination with $Hantavax^{TM}$. Seroconversion rates were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA), and plaque reduction neutralization test (PRNT). Seroconversion rates of 21 vaccinees at one year after primary basic vaccination were 52.3%, 95.2%, 0.0%, 47.6%, and 28.6%, and 13 vaccinees at one month after 1st booster vaccination were 100%, 100%, 30.7%, 100% and 100% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates declined slightly by twenty months, and they were 84.6%, 92.3%, 0.0%, 84.6% and 69.2% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates of 9 vaccinees at three months after 2nd booster vaccination were 100%, 100%, 0.0%, 100%, and 88.9%, and 16 vaccinees at one year after the 2nd booster vaccination were 87.5%, 93.8%, 0.0%, 87.5% and 81.3% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Based on the above result $Hantavax^{TM}$ has proved a vigorous anamnestic response after the 1st and the 2nd booster vaccination and has persisted higher fluorescence, agglutination and neutralizing antibody titers in vaccinees.

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Effect of a trivalent (FPV, FHV, FCV) inactivated vaccine in kittens (고양이 3종(FPV, FHV, FCV) 불활화 백신의 효과)

  • Lee, Sung-min;Yoon, In-joong;Choi, Hwan-won;Lee, Keun-jwa;Lee, Kyoung-youl;Kim, Moo-kang
    • Korean Journal of Veterinary Research
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    • v.45 no.3
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    • pp.311-323
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    • 2005
  • This study tested the effect of a trivalent (feline panleukopenia; FPV, feline viral rhinotracheitis; FHV, feline calicivirus infection; FCV) inactivated vaccine in cats. The vaccine was tested for the safety in guinea pigs, mice and cats. Also, it was tested for the efficacy in cats. The vaccine was inoculated to cats at 7~9 and 10~12 weeks of age (conventional schedule) and the serological response to vaccination was assessed and was compared to the unvaccinated group. All cats were bled by jugular venipuncture for FPV, FHV and FCV specific serological test (virus neutralizing antibody, VN) at 7~9, 10~12 and 13~15 weeks. After last bleeding, all cats were inoculated with each virus (FPV : orally $2ml\;10^{7.5}\;TCID_{50}/ml$, FHV : nasally $1ml\;10^{7.0}\;TCID_{50}/ml$ and FCV : nasally $1ml\;10^{7.0}\;TCID_{50}/ml$). The Vaccine verified excellent protective effect in guinea pigs, mice and cats. The VN antibody titers of the unvaccinated group cats against FPV, FHV and FCV were <2~16, on the other hand the vaccinated group cats were $512{\sim}{\geq}4096$, 64~1024 and 64~1024, respectively. When all cats were challenged with virulent viruses, the survival rates of the vaccinated group cats were over 80%, while the survival rates of the unvaccinated group cats were less 20%. The typical clinical signs were not observed in the vaccinated group cats, but the typical clinical signs and histopathological lesions were observed in the unvaccinated group cats. As the result of tests, the VN values obtained in this study appeared to be high enough to protect cats from viral challenges. The trivalent (FPV, FHV, and FCV) inactivated vaccine seemed to be very effective, for prevention of feline viral diseases (FPV, FHV, and FCV).