• Title/Summary/Keyword: virus titer

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Expression and Immunogenicity of SARS-CoV-2 Virus-Like Particles based on Recombinant Truncated HEV-3 ORF2 Capsid Protein

  • Zhou, Yong-Fei;Nie, Jiao-Jiao;Shi, Chao;Ning, Ke;Cao, Yu-Feng;Xie, Yanbo;Xiang, Hongyu;Xie, Qiuhong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.10
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    • pp.1335-1343
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    • 2022
  • COVID-19 is an emerging disease that poses a severe threat to global public health. As such, there is an urgent demand for vaccines against SARS-CoV-2, the virus that causes COVID-19. Here, we describe a virus-like nanoparticle candidate vaccine against SARS-CoV-2 produced by an E. coli expression system. The fusion protein of a truncated ORF2-encoded protein of aa 439~608 (p170) from hepatitis E virus CCJD-517 and the receptor-binding domain of the spike protein from SARS-CoV-2 were expressed, purified and characterized. The antigenicity and immunogenicity of p170-RBD were evaluated in vitro and in Kunming mice. Our investigation revealed that p170-RBD self-assembled into approximately 24 nm virus-like particles, which could bind to serum from vaccinated people (p < 0.001) and receptors on cells. Immunization with p170-RBD induced the titer of IgG antibody vaccine increased from 14 days post-immunization and was significantly enhanced after a booster immunization at 28 dpi, ultimately reaching a peak level on 42 dpi with a titer of 4.97 log10. Pseudovirus neutralization tests showed that the candidate vaccine induced a strong neutralizing antibody response in mice. In this research, we demonstrated that p170-RBD possesses strong antigenicity and immunogenicity and could be a potential candidate for use in future SARS-CoV-2 vaccine development.

Changes of maternal antibodies in chicks vaccinated to breeder against infectious bronchitis, infectious Bursal disease, and Newcastle disease virus (모계의 전염성기관지염, 전염성 F낭병 및 뉴캣슬병 백신투여에 따른 모체이행 항체의 변동)

  • 고원석;김태중;이정원;서이원;송희종;오언평
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.133-139
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    • 1998
  • Serum samples collected from 30 breeders and their progeny 30 chicks. The antibodies against infectious bronchitis(IB), infectious bursal disease (IBD) and Newcastle disease(ND) viruses were detected by ELISA using commercial ELISA kit. The breeders were vaccinated against IB, IBD and ND viruses according to general vaccination program. Geometric mean titers(GMT) of ELISA were monitored from 1-day old to 17-day old chicks and compared with breeder chickens. The GMT of ELISA to IB, IBD and ND were declined half level of the breeder antibody titer at 6-, 8- and 7-day old. And, the GMT of ELISA to IB, IBD and ND were declined than that of protective titer at 6-, 1-, and 4-day old. Thereafter, the GMT of ELISA was declined and disappeared according to ages of chicks. Taken together, this study led to conclusion that time-course of maternal antibody titers of chicks from vaccinated breeders, and this is very important data for vaccination to chicks.

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Comparison of pooled Versus Individual Sera in Avian Infectious Bronchitis Virus Seroprevalence Study (닭 전염성 기관지염 바이러스의 혈청 유병률 연구에서 개별혈청과 합병혈청의 비교)

  • Kim, Sa-Rim;Kwon, Hyuk-Moo;Sung, Haan-Woo;Pak, Son-Il
    • Journal of Veterinary Clinics
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    • v.23 no.4
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    • pp.416-420
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    • 2006
  • Compare to testing sera individually, pooled-serum testing has considered as a cost-effective method, particularly on a large population-based seroprevalence studies. This study was to determine the relationship between individual sera and pooled sera titers for detection of avian infectious bronchitis virus (IBV) and to evaluate suitability of pooled sera by comparing prevalences estimated from both samples. A total of 5,000 individual samples were collected from 500 flocks in Chungcheong, Gyunsgi, and Kangwon provinces between January 2005 and February 2006. Ten samples were randomly selected from each flock. Five-hundred pooled sera were prepared by mixing equal amount of each 10 individual serum from the original samples. IBV antibody titers were measured by hemagglutination inhibition (HI) test. The least squares regression analysis was performed to construct equation between pooled and mean individual titers. To determine whether the flock is infected 4 arbitrary criteria were used: detection of at least 1 chicken with HI titer ${\ge}$ 9 (criterion 1), detection of at least 2 samples with HI titer ${\ge}$9 (criterion 2), detection of at least 1 sample with HI titer ${\ge}$ 10 (criterion 3), and filially detection of at least 1 sample with HI titer ${\ge}$ 11 (criterion 4). The receiver operating characteristic (ROC) curve was used to examine the cut-off points of pooled titers showing optimal diagnostic accuracy. The area under the curve (AUC), sensitivities (Se), specificities (Sp), and positive (PPV) and negative (NPV) predictive values were calculated. The regression equation between pooled titers (pool) and mean individual titers (mean) was: $pool= 1.2498+0.8952{\times}mean$, with coefficient of determination of 87% (p< 0.0001). The optimal cut-off points of pooled titers were titer 8 for criterion 1 (AUC=0.975, Se=0.883, Sp=0.959, PPV=0.985, NPV=0.728), titer 8 for criterion 2 (AUC=0.969, Se=0.954, Sp=0.855, PPV=0.926, NPV=0.907), titer 9 for criterion 3 (AUC=0.970, Se=0.836, Sp=0.967, PPV=0.978, NPV=0.772), and titer 9 for criterion 4 (AUC= 0.946, Se=0.928, Sp=0.843, PPV=0.857, NPV=0.921). The difference of 'prevalence estimated by individual and pooled sample showed a minimum of 2% for criteria 2 and a maximum of 9.1:% for criteria 3. These results indicate that the use of pooled sera in HI test for screening IBV infection in laying hen flocks is considered as a cost-effective method of testing large numbers of samples with high diagnostic accuracy.

Mode of Antiviral Activity of Water Soluble Components Isolated from Elfvingia applanata on Vesicular Stomatitis Virus

  • Eo, Seong-Kug;Kim, Young-So;Oh, Ki-Wan;Lee, Chong-Kil;Lee, Young-Nam;Han, Seong-Sun
    • Archives of Pharmacal Research
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    • v.24 no.1
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    • pp.74-78
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    • 2001
  • A preparation of water soluble components (EA) was made from carpophores of Elfvingia applanata (Pers.) Karst and its in vitro antiviral activity on vesicular stomatitis virus [(Indiana serotype, VSV(IND)] was investigated by plaque reduction assay. EA exhibited potent antiviral activity on VSV(IND) growth and negligible cytotoxicity on Vero cells, 50% effective concentration ($EC_{50}C$/) of 104$ug\textrm\/ml$ and 50% cytotoxic concentration ($CC_{50}C$) of 3,793$ug\textrm\/ml$, respectively. Selectivity index (Sl $CC_{50}C$/$EC_{50}C$) of EA on Vero cell and VSV(IND) was about 36.5. EA did not display either a direct virucidal effect on V5V(IND) or induction of antiviral substance by Vero cells upon its treatment. Thus, the mode of antiviral activity of EA was studied at steps of viral adsorption onto cell. When both EA and virus were added to cell monolayers, titer of cell-free virus in culture supernatant increased in ca. 30-40% compared with that of control group and titer of cell-associated virus was 60-100% higher than that of control group. These results suggested that antiviral activity of EA on VSV(IND) might be due to the hindrance of viral entry to cells at eITher endocytosis or loss of envelope.

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Solvent/Detergent Inactivation and Chromatographic Removal of Human Immunodeficiency Virus During the manufacturing of a High Purity Antihemophilic Factor VIII concentrate

  • Kim, In-Seop;Park, Yong-Woon;Woo, Hang-Sang;Chong E. Chang;Lee, Soungmin
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.187-191
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    • 2000
  • A validation study was conducted to determine the efficacy of solvent/Detergent (S/D) inactivation and Q-Sepharose column chromatographic removal of the human immunodeficiency virus (HIV) during the manufacturing of a high purity antihemopilic factor VIII (GreenMono) from human plasma. S/D treatment using the organic solvent, tri (n-butyl) phosphate, and the detergent, Trition X-100, was a robust and effective step in eliminating HIV-1. The HIV-1 titer was reduced from an initial titer of 8.3 log10 TCID50 to undetectable levels within one minute of S/D treatment, HIV-1 was effectively partitioned form factor VIII during Q-Sepharose column chromatography with the log reduction factor of 4.1 . These results strongly assure the safety of GreenMono From HIV.

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Prevalence on protective serum antibodies of canine distemper virus and canine parvovirus in Ulsan area (울산지역의 개 디스템퍼 및 파보 장염의 항체보유 실태 조사)

  • Sung, Ki-Chang;Lee, Eun-Woo;Park, Chang-Eun
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.213-221
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    • 2010
  • The results from a total of 412 blood samples consisted of 187 samples from regular visiting group (RV), 94 samples from first visiting group (FV), 52 samples from abandoned group (A), 54 samples from special breeder group (SB), and 25 samples from preliminary breeder group (PB) showed that RV(94.7%) and SB(88.9%) groups had the higher levels of protective antibody, but PB (36.0%) group revealed the lowest level. Among 96 blood samples with lower protective antibody levels, 14 samples (14.6%), 72 samples (75.0%) and 10 samples (10.4%) were below the protective antibody levels to distemper/parvo-virus, distemper only and parvovirus only, respectively. These results implied that antibody to parvovirus was well generated than that to distemper. Eighty six samples (20.9%) showed the protective antibody titer under 1:96 to distemper and 24 samples (5.8%), the protective antibody titer under 1:40 to parvovirus.

Studies on the immunization against field strain after live Newcastle disease virus vaccination (뉴캣슬병 생독백신 접종 후 야외 분리 바이러스에 대한 면역성 조사)

  • 김순태;박인화;김성국;김영환;조광현;손재권
    • Korean Journal of Veterinary Service
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    • v.24 no.2
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    • pp.147-159
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    • 2001
  • This Study was conducted to determine vaccination programs for the control of Newcastle Disease(ND) in chickens and investigate protective effect against Newcastle disease virus (NDV) after live ND vaccination. Maternal HI antibody titer level of chickens according to day(age) 1, 7, 14, 21, 28 and 35 were decreased gradually as 7.10$\pm$0.74, 6.57$\pm$0.74, 3.71$\pm$1.25, 2.20$\pm$1.03, 1.20$\pm$1.23 and 0.50$\pm$0.71. As a result of HI test and ELISA, both chickens vaccinated with VG/GA strain live vaccine at 1-day-old and chickens not vaccinated do not have antibody titer for protection against NDV at 14-day-old. Except for LaSota strain vaccine, in case of vaccination with VG/GA spray and VG/GA, B1 and LaSota strain drinking water at 14-day-old, the protective effect was 100% in chickens inoculated NDV($10^{7.2}$ $EID_{50}$/50${\mu}\ell$, eye drop) at 21-day-old, but not 10~50% at 28-day-old. These data suggest that live NDV vaccination should be given at 10-day-old 20-25day-old for protect against NDV at periodic outbreaks of ND caused by velogenic viscerotropic NDV in the environment of a farm.

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A survey of newcastle disease virus antibody titers on slaughtered broiler chicks (도축 육계에서 뉴캣슬형 바이러스에 대한 혈중항체가 조사)

  • 이정원;허철호;이종환;권정택;송희종
    • Korean Journal of Veterinary Service
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    • v.24 no.3
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    • pp.217-222
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    • 2001
  • Hemagglutination inhibition(HI) titers of Newcastle disease(ND) were measured to investigate the vaccination times on three different species of broiler chicks in Chonbuk province. Each 330 of Cobb, Ross and White-semi broiler chicks were selected from 11 broiler farms. The primary vaccine were sprayed in hatchery at one day old chicks. Secondary and tertiary vaccine were used by drinking water at 7 to 24 days old chicks. The ND antibody titer were measured by HI from each different species of broiler chicks at the marketing date. Total average HI titers of Cobbs vaccinated with primary ones, secondary and tertiary ones were recorded 1.86, 1.52 and 2.76, respectively. The antibody titers were shown to 2.22, 2.13, 3.07 in terms of vaccination of Ross broiler chicks. They were also 2.56, 2.65 and 2.78 in terms of vaccination of White-semi broiler chicks. The value HI titer were not statistically different of all treatments. The results of this experiment suggested that HI titer of sera is scored less than defensive value of ND antibody titer at more than two times of vaccination.

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Isolation and cultivation of swine encephalomyocarditis virus (돼지 뇌심근염 바이러스의 분리 배양)

  • Ha, Yong-kong;Yoon, Seok-min;Jung, Byung-tack;Park, Nam-yong;Lee, Bong-ju;Chung, Chi-young;Kee, Hye-young;Bae, Seong-yeol
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.479-484
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    • 1991
  • Encephalomyocarditis(EMC) virus was isolated from the mummified and stillborn pigs at a swine farm in Chonnam Province, experienced with EMC infection over the period Oct.~Dec. of 1989. In addition some cultural, serological properties of the isolates and experimental infections in the piglets were studied. The results obtained were as follows; 1. Two EMC virus strains with HA titers and CPE similar to EMC-ATCC were established in a baby hamster kidney (BHK)-21 cell line by inoculating homogenates of brain and heart of the 19 mummified or stillborn pigs and designated $K_3$ and $K_{11}$. 2. At the second BHK-21 cell line passage of the initial isolates CPE appeared after incubation for 16~18 hours, while at the fourth and fifth passage the highest titer of HA was recorded, titer of HA using rat and guinea pig erythrocytes. 3. One pig inoculated with the isolate $K_3$ showed dyspnea as clinical signs and died at the 10 days after inoculation at necropsy white necrotic foci were observed from the dead animal heart. 4. Although all the rest surviving pigs showed increases in antibody titer and body temperature of $40^{\circ}C$ above for the initial 2~4 days followed by the return to normal, there were no gross lesions when the animals were sacrificed at the 2 weeks after inoculation.

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Effect of Dietary Vitamin E on Growth Performance and Immune Response of Breeder Chickens

  • Lin, Y.F.;Chang, S.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.884-891
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    • 2006
  • The effect of dietary vitamin E supplementation on immune responses was studied in breeder chickens during the maturing period. In experiment 1, 17-week old female birds were fed corn-soybean meal based diets supplemented with either 0, 40, 80, 120, or 160 mg vitamin E (all-rac-${\alpha}$-tocopherol acetate)/kg diet for 19 weeks. In experiment 2, 23-week old male birds were fed the corn-soybean meal based diet supplemented with either 0, 20, 40, 80 or 160 mg vitamin E/kg diet for 8 weeks. The chickens were evaluated for growth performance, antibody titer to sheep red blood cell (SRBC), Newcastle disease virus (NDV), infectious bursal disease virus (IBDV) and infectious bronchitis virus (IBV), and skin response to phytohemagglutinin-P (PHA-P). The results showed that supplemental vitamin E improved body weigh gain of laying pullets during peak-laying period but had no significant effect on growth performance of cockerels. For cockerels, addition of 20 mg vitamin E/kg diet significantly enhanced (p<0.05) immune response to SRBC compared to those added with 0, 80 and 160 mg vitamin E/kg diet; addition of 20 mg vitamin E/kg diet had higher (p<0.01) antibody titer to IBDV than those added with 40-160 mg vitamin E/kg diet. No significant effects on immune response were observed in laying pullets fed supplemental vitamin E. The findings suggest that moderate supplementation of vitamin E may enhance immune responses to selective antigens in cockerels but excessive vitamin E may depress specific immune response.