• Title/Summary/Keyword: virus titer

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Detection of antibodies in swine serum to Aujeszky's disease virus using agar-gel immunodiffusion test (Agar-gel immunodiffusion test를 이용한 돼지 혈청중 Aujeszky's disease virus 항체 검출에 관한 연구)

  • Cho, Hyo-gueon;Jun, Moo-hyung
    • Korean Journal of Veterinary Research
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    • v.30 no.3
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    • pp.297-307
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    • 1990
  • To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

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Immune response and efficacy of pigeon pox virus vaccine and fowl pox virus vaccine in chickens

  • Shil, Niraj Kanti;Rahman, Md Siddique;Hossain, M.T.;Islam, M.T.;Rahman, M.M.;Islam, K.B.M. Saiful;Cha, Se-Yeoun;Jang, Hyung-Kwan;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.497-503
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    • 2007
  • The humoral immune response of chicken vaccinated with fowl and pigeon pox virus vaccines was determined with the protective potentiality of the two vaccines in field condition of Bangladesh. Different aged Fayoumi chicks were subjected for the study. To assess the relationship with better immune response among experimental groups, the average percentage of 'take reaction' was examined and recorded to 97.77% in group A, 93.33% in group B and 100.0% in group C. The level of immune status induced by different vaccinated group was measured by passive hemagglutination (PHA) microplate test method. The mean PHA titer levels after primary vaccination were $33.06{\pm}14.13$ in group A, $32.0{\pm}14.81$ in group B, and $33.0{\pm}13.66$ in group C. Following booster vaccination, the mean PHA titer levels in prior of challenge were increased to $55.46{\pm}14.64$ in groups A and C, and $46.93{\pm}16.52$ in group B. The recorded PHA titer levels of each group at two weeks after challenge were significantly increased to $106.66{\pm}31.22$, $93.86{\pm}33.04$ and $110.93{\pm}29.29$, respectively. The PHA titer levels after vaccination and challenge were significantly increased compared to pre-vaccination titer levels (P<0.01). Although the PHA titer levels among three groups administrated different vaccine combinations in prior of challenge were significantly varied (P<0.01), it was observed that all of the vaccinated chicks were highly protected against challenge infection.

Comparative study on avian influenza virus antibody titer by hemagglutination inhibition test and enzyme-linked immunosorbent assay in the mass zone layer (산란계 밀집지역에서 혈구응집억제반응과 효소면역측정법을 이용한 가금인플루엔자 혈중항체가의 비교 조사)

  • 이정원;엄성심;이성재;서이원;서석열;정동석;송희종
    • Korean Journal of Veterinary Service
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    • v.27 no.1
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    • pp.81-87
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    • 2004
  • This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA), the HI titer and mean ratio S/P ratio) of avian influenza virus. To perform this study, the 1,457 sera of layers 21 farms in May, July and September, respectively. As a result of HI test, positive rates were 480 to 422 (92.1%) in May, 494 to 394(79.8%) in July and 483 to 402(83.2%) in September, and the mean antibody titer were 4.6, 4.3, 4.0 to 0.3 decreased, respectively. The positive rates by ELISA, 480 to 475(99.0%) in May, 494 to 485(98.2%) in July, 483 to 472(97.7%) in September, and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. The result of HI test and ELISA positive 480 to 422(92.1%), 475(99.0%), 494 to 394(79.8%), 485(98.2%) and 483 to 402(83.2%), 472(97.7%). Therefore, ELISA was shown more sensitive compare the HI titers.

Correlations in the results of virus neutralization test, hemagglutination inhibition test, and enzyme-linked immunosorbent assay to determine infectious bronchitis virus vaccine potency

  • Park, Mi-Ja;Joh, Seong-Joon;Choi, Kang-Seuk;Kim, Aeran;Seo, Min-Goo;Song, Jae-Young;Yun, Seon-Jong
    • Korean Journal of Veterinary Research
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    • v.56 no.3
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    • pp.189-192
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    • 2016
  • The virus neutralization (VN) test was used to determine potency of the infectious bronchitis (IB) vaccine. The results of VN, hemagglutination inhibition (HI), and enzyme-linked immunosorbent assay (ELISA) were compared with those of the IBV M41. The $r^2$ values between VN and HI titers and the ELISA antibody titer were 0.8782 and 0.0336, respectively, indicating a high correlation between VN and HI, but not VN and ELISA. The Cohen's kappa coefficient between the VN titer of 2 $log_{10}$ and HI titer of 5 $log_2$ was 0.909. Our results showed that VN could be replaced with HI for testing the potency of IBV M41.

Investigation of hematological analyses and major viral serum antibody titers in DongGyeongi (동경이의 혈액상 및 주요 바이러스 혈중 항체가 조사)

  • Choi, Seog-Gyu;Sung, Gi-Chang;Lee, Eun-Woo;Park, Chang-Eun
    • Korean Journal of Veterinary Service
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    • v.37 no.4
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    • pp.273-280
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    • 2014
  • Investigations of hematologic and canine distemper virus, parvovirus antibody titer for DongGyeongi were performed. This study was conducted to determine into feeding and management blood values on DongGyeongi. Blood samples were collected from 110 healthy dogs (male 60, female 50). The diagnostic virus disease and hematologic results were classified by age, sex, color. Although gender differences were not apparent, complete blood cell counts analyses were performed and analyzed accordingly. Statistically significant differences (P<0.05) specific to age (<2). Also, The canine distemper virus and parvovirus antibody titer were correlation to age dependent (P<0.05). In conclusion, data obtained from this study may be valuable as a standard for interpretation of the results in hematologic and major viral antibody titer analysis of DongGyeongi populations. For the management of DongGyeongi, the programs will be used for the epidemic disease prevention.

Large-Scale Culture of Hepatitis A Virus in Human Diploid MRC-5 Cells and Partial Purification of the Viral Antigen for Use as a Vaccine

  • Kim, Hyun-Seok;Chung, Yong-Ju;Jeon, Yeong-Joong;Lee, Sung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.386-392
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    • 1999
  • A large-scale culture of hepatitis A virus in human diploid MRC-5 cells was conducted. In a roller bottle culture, the virus was grown to a maximum titer in 3 weeks after infection. Over 95% of the cell-associated virus was excreted after culturing the infected cells in suspension media without fetal bovine serum for 3 days. The cultured virus was inactivated with formalin, concentrated by ultrafiltration, and partially purified by ultracentrifugation in a non-ionic gradient medium of Renocal. Two separate peak fractions showing high anti-HAY ELISA titer were pooled and about 40% of HAV antigen was recovered by this purification procedure. Of the partially purified vaccine, the protein pattern in SDS-PAGE and immunogenicity in mice were compared with a commercial HAV vaccine. In SDS-PAGE, the purified vaccine in this study and the commercial vaccine showed almost the same protein pattern. The seroconversion rate of the purified vaccine in mice was not different from that of the commercial vaccine. Therefore, we could prepare a good grade of HAV vaccine by a simple purification procedure although the purification itself was not completed.

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Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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A sere-epidemiological survey on Ibaraki disease in western area of Kyunggi province (경기 서부지방의 소 Ibaraki병 중화항체가 조사)

  • 이우종;고신일;최영래;강영배;최강석
    • Korean Journal of Veterinary Service
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    • v.19 no.3
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    • pp.207-211
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    • 1996
  • To investigate the serum neutralizing antibodies against Ibaraki disease virus in the western area of Kyunggi province, a sero-epidemiological survey was done from August 1995 to March 1996. The results obtained are summarized as follows : 1. An overall prevalence of the neutralizing antibodies agaist Ibaraki virus was as high as 68.6% (218 positive reactors amomg 318 heads of dairy cattle). 2. It showed the regional differences with 60.5${\ulcorner}$(46/76) in Koyang, 75.2% (100/133) in Paju and 66.1% (72/109) in Kimpo. 3. It also appeared with a seasonal difference showing 74.4% of prevalence with the mean titer higher than 60 during the mosquito season (from August to November) and 58.6% of prevalence with the mean titer 22 after the mosquito season to March. 4. Any cross reactions between Ibaraki and bluetongue viruses were not detected in the ELISA and AGID tests.

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Strategies for Transmissible Gastroenteritis Virus(TGEV) Vaccine Production by Swine testicle cells

  • Kim, Min-Young;Jeong, Yeon-Ho;Kim, Myoung-Hwa;Ko, Yun-Mi
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.203-206
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    • 2005
  • TGE (Transmissible Gastroenteritis) caused by a virus belonging to family coronavirus, results in an acute infection of the small intestine of the pig. The optimum operation variables such as multiplicity of infection (MOI), infection time and harvest time were investigated for TGE vaccine production by immobilized ST(swine testicle) cells. In the culture supplemented with 5% serum, maximum virus titer of $1.2{\times}10^6pfu/ml$ was obtained at the conditions of 0.01 MOI, 2day infection time, and 1 day harvest time. Serum is a potential source of bacterial, mycoplasmal and viral contamination, and it has a possibility of the introduction of serum proteins, prion and pyrogens into the final product. For these reasons, much attention has been focused on the development of serum-free media. A new serum-free media (SFM) has been developed in order to produce TGE vaccine of high quality with low cost. The performance of SFM developed was compared with other commercially available serum-free media and serum supplemented media in terms of virus productivity. The cultures with serum-free media showed higher titer than that with serum supplemented media. Among various serum-free media tested, CHO-S-SFMII showed highest virus titer.

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Experimental infection in guinea pig with foot and mouth disease virus

  • Abdul-Ahad;Rahman, Md-Siddiqur;Rahman, Md-Mostafizur;Baek, Byeong-Kirl;Lee, John-Hwa
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.73-80
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    • 2003
  • In order to obtain information on murine model for foot and mouth disease virus(FMDV) type Asia 1, we studied whether guinea pig was a suitable model for studying FMDV. Apparently healthy 3 months old albino guinea pigs and unweaned 3 days old Swiss albino mice were used for this study. Total of 8 guinea pigs were divided into the infected(n=5) and control(n=3) groups. The incubation period of FMDV in the guinea pigs were roughly 2 days and the viremia persisted for 3 days in the guinea pigs. Mice inoculated with the plasma from control guinea pigs did not show any sign of viremia. The plasma were titrated by virus neutralization test using suckling mice as an indicator host. The mean virus neutralizing antibody titers of infected guinea pig at 3 DPI, 4 DPI and 5 DPI were log$\_$10/2.16, log$\_$10/ 3.39 and log$\_$10/ 3.44, respectively whereas there was no neutralizing antibody titer in control group. The difference between the mortality pattern and mean virus neutralizing antibody titer of infected and that of control group at day 3, 4, 5 were statistically significant(p<0.0l).