• Title/Summary/Keyword: virus titer

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Effects of fetal bovine serum concentrations on viral infectivity titers of infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus (Fetal bovine serum의 농도에 따른 infectious hematopoietic necrosis virus와 infectious pancreatic necrosis virus의 감염가 변화)

  • Kim, Hyoung Jun;Park, Jeong Su;Kwon, Se Ryun
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.81-85
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    • 2018
  • Fetal bovine serum (FBS) is an essential element of cell growth and can also affect the viral replication. In this study, we tried to find out whether FBS concentration affects the viral infectivity titer of IHNV and IPNV. EPC cells were suspended with MEM supplemented with various concentrations of FBS (MEM0, MEM2, MEM5 and MEM10) and cultured in 96-well plate. Each virus was 10-fold diluted virus and inoculated in 96-well plate. The highest infectivity titer of IHNV was $10^{7.88}\;TCID_{50}/mL$ in 96-well plate using MEM5 and the lowest one was $10^{7.30}\;TCID_{50}/mL$ in 96-well plate using MEM10. The highest infectivity titer of IPNV was $10^{7.47}\;TCID_{50}/mL$ in 96-well plate using MEM5 and the lowest one was $10^{6.97}\;TCID_{50}/mL$ in 96-well plate using MEM10. This study showed that not only 0% FBS but 10% FBS leads low infectivity titer of IHNV and IPNV. Therefore, it is considered that the desirable concentration of FBS is 2% or 5% for measurement of infectivity titer of IHNV and IPNV.

Identification of Endogenous Genes for Normalizing Titer Variation of Citrus Tristeza Virus in Aphids at Different Post-acquisition Feeding Times

  • Wang, Hongsu;Chen, Qi;Liu, Luqin;Zhou, Yan;Wang, Huanhuan;Li, Zhongan;Liu, Jinxiang
    • The Plant Pathology Journal
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    • v.38 no.4
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    • pp.287-295
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    • 2022
  • Citrus tristeza virus (CTV) is efficiently transmitted in a semi-persistent manner by the brown citrus aphid (Toxoptera citricida (Kirkaldy)). Currently, the most sensitive method for detecting plant viruses in insect vectors is reverse-transcription quantitative polymerase chain reaction (RT-qPCR). In this study, the elongation factor-1 alpha (EF-1α) gene and acidic p0 ribosomal protein (RPAP0) gene were confirmed to be suitable reference genes for RT-qPCR normalization in viruliferous T. citricida aphids using the geNorm, NormFinder, and BestKeeper tools. Then the relative CTV titer in aphids (T. citricida) at different post-acquisition feeding times on healthy plants was quantified by RT-qPCR using EF-1α and RPAP0 as reference genes. The relative CTV titer retained in the aphids gradually decreased with increasing feeding time. During the first 0.5 h of feeding time on healthy plants, the remaining CTV titer in aphids showed about 80% rapid loss for the highly transmissible isolate CT11A and 40% loss for the poorly transmissible isolate CTLJ. The relative CTV titer in aphids during more than 12 h post-acquisition times for CT11A was significantly lower than at the other feeding times, which is similar to the trend found for CTLJ. To our knowledge, this is the first report about the relative titer variation of CTV remaining in T. citricida at different post-acquisition feeding times on healthy plants.

Effect of Formalin Inactivation on Viral Hemorrhagic Septicemia Virus (VHSV) (Viral Hemorrhagic Septicemia Virus (VHSV)에 대한 포르말린 불활화 의 영향)

  • Park, Jeong Su;Kim, Hyoung Jun;Joo, Young Hun;Kwon, Se Ryun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.6
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    • pp.644-649
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    • 2019
  • Killed vaccines, developed by inactivation with formalin, have been investigated for many fish viruses. In this study, the inactivation of viral hemorrhagic septicemia virus (VHSV) by formalin was investigated based on the infectivity titer. When viral cell culture supernatants were used, the infectivity titer decreased 1,000-fold at 1 d after treatment with 0.1% (v/v) formalin, but was below the detection limit at 7 and 14 d. Moreover, neither the N nor G gene were detectable by RT-PCR immediately after formalin treatment. In western blot analysis, N protein was not detected by rabbit antiserum against VHSV KR-9225 from 2 d after formalin treatment. On the other hand, when we used a virus that was purified and concentrated ~100 times, the infectivity titer was maintained at 106.05 TCID50/mL, even at 14 d after formalin treatment, and no change in the viral structural proteins was observed. This study provides important data on the production and use of formalin-inactivated vaccines.

Hemagglutination of Hantaan and Seoul Viruses (한탄바이러스와 서울바이러스의 혈구응집반응)

  • Seong, In-Wha;Song, Ki-Joon;Lee, Ho-Wang
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.2
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    • pp.227-231
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    • 1986
  • The hemagglutination activities of Hantaan virus and Seoul virus were demonstrated. The hemagglutinins were prepared by sucrose-acetone extraction method from suckling mouse brains infectecd with Hantaan and Seoul viruses. Hemagglutination of goose erythrocytes by these viral hemagglutinins was pH dependent in phosphate buffer system. Hantaan and Seoul viruses were distinguished by pH range of hemagglutination. 76/118 and 79/90 strains of Hantaan virus showed hemagglutination at the range of pH 5.75-6.4 and the optimal pH was 5.75 with the titer of 1:512 in 76/118 and 1:256 in 79/90. In contrast, KSNUSD 84/34 strain of Seoul virus revealed hemagglutination at the range of pH 6.2-6.4 and the optimal pH was 6.4 with the titer of 1 : 64.

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Effect of Modified Live Virus Vaccine aganist Canine Parvovirus Infection (개 파보바이러스 감염에 대한 Modified Live Virus Vaccine의 효과)

  • 한정희;유기일;권혁무;서강문
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.46-55
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    • 1998
  • This study was performed to verify the effect of modified live virus vaccine against canine parvovirus (CPV) infection. Serum hemagglutination inhibition (Hl) test, histopathological and immunohistochemical techniques and polymerase chain reaction were used. The results were as follows: 1. During the experimental terms after vaccination, serum Hl titer was stable. Geometric mean titer (GMT) after the 1st vaccination was 280. After virulent CPV was challenged, GMT was 1,306. 2. After challenge by virulent CPV, the vaccinated group was not shown clinical signs and gross and histopathological findings. 3. After challenge by virulent CPV, the vaccinated group was not detected viral antigens in the small intestine immunohistochemically. 4. After challenge by violent CPV, the vaccinated group was not shown virus shedding in feces. In conclusions the overall results confirmed that modified live virus vaccine was effective on prevention of canine parvovirus infection.

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Survey of foot-and-mouth disease virus structural protein antibody titer in Yeongcheon (영천지역 구제역 바이러스 구조단백질 항체가 조사)

  • Sohn, Jun-Hyung;Hwang, You-Sun;Sohn, Kyu-Hee;Shin, Sung-Ho;Lee, Eun-Mi;Kim, Soon-Tae;Cho, Min-Hee;Yun, Mun-Jo
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.13-17
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    • 2015
  • Three serotypes (O, A and Asia1) of the foot-and-mouth disease (FMD) vaccine were injected into domestic cloven-hoofed animals in korea after the nationwide spread at the end of 2010. The purpose of this study was survey of FMD virus stuructural protein (SP) antibody titer in Yeongcheon by enzyme-linked immunosorbent assay (ELISA). Total 1,324 samples collected from 89 farms were tested. The overall seroprevalence of FMD virus SP antibodies was 58.8% (778/1,324) The seroprevalence of FMD virus SP antibody varied with species. Results in cattle (over 12 month old) and pig (90 to 130 day old) were 58.8% and 44.9% respectively.

Expression of the E. coli LacZ Gene in Chicken Embryos Using Replication Defective Retroviral Vectors Packaged With Vesicular Stomatitis Virus G Glycoprotein Envelopes

  • Kim, Teoan;Lee, Young Man;Lee, Hoon Taek;Heo, Young Tae;Yom, Heng-Cherl;Kwon, Mo Sun;Koo, Bon Chul;Whang, Key;Roh, Kwang Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.163-169
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    • 2001
  • Despite the high potency of the retrovirus vector system in gene transfer, one of the main drawbacks of has been difficulty in preparing highly concentrated virus stock. Numerous efforts to boost the virus titer have ended in unsatisfactory results mainly due to fragile property of retrovirus envelope protein. In this study, to overcome this problem, we constructed our own retrovirus vector system producing vector viruses encapsulated with VSV-G (vesicular stomatitis virus G glycoprotein). Concentration process of the virus stock by ultracentrifuge did not sacrifice the virus infectivity, resulting in more than 108 to 109 CFU (colony forming unit) per ml on most of the target cell lines tested. Application of this high-titer retrovirus vector system was tested on chicken embryos. Injection of virus stock beneath the blastoderms of pre-incubated fertilized eggs resulted in chick embryos expressing E. coli LacZ gene with 100% efficiency. Therefore, our results suggest that it is possible to transfer the foreign gene into chicken embryo using our high-titer retrovirus vector.

A survey of Newcastle disease virus antibody titers on slaughtered chickens (도계장 출하 닭에 대한 뉴캐슬병 항체가 조사)

  • Heo Jung-Ho;Lee Kuk-Cheon;Cho Myung-Heui;Kim Kuk-Hun;Hah Dae-Sik;Kim Jong-Shu
    • Korean Journal of Veterinary Service
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    • v.29 no.3
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    • pp.267-276
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    • 2006
  • To investigate serum antibody titers of Newcastle disease (ND) virus, serum samples were collected from 19,774 slaughtered chickens from 862 farms in Geoje, Goseong, Tongyung and other province during the period from January 2001 to December 2005. The results obtained are summarized as follows; 1. The negative percentage of ND antibody titer with below 1.0 levels decreased from 46.7% in 2001 to 40.5% in 2004 but those of it increased to 47.0% in 2005 and the positive percentage of ND antibody titer with higher 5 levels increased from 15.8% in 2001 to 26.2% in 2005. 2. The season levels of ND antibody titer showed various levels according to seasons but the general trend of ND antibody titer decreased in the later half of the year than those of the first half of the year. 3. The negative percentage of ND antibody titer on provinces decreased from 52.0% (2001) to 36.2% (2004) in Goseong but those of it increased to 51.4% in 2005. The negative percentage of ND antibody titer of Tongyung and other provinces increased from 51.0%, 28.5% (2001) to 55.0% and 44.3% (2005), respectively. 4. The year average levels of ND antibody titer increased from 2.2 to 2.6 during 2001- 2004 but those of it decreased to 2.4 in 2005. The provinces average levels of ND antibody titer increased from 2.0 to 2.8 during 2001 - 2005 and others provinces except Gyeongnam decreased from 3.3 to 2.6 during the 2001-2005. 5. The farm average levels of ND antibody titer with below 1.0 levels decreased from 26.3% in 2001 to 10.2% in 2004 but those of it increased to 17.6% in 2005 and those of below 1.0 or 2.0 levels increased from 17.9% (2001) to 27.7% (2005). The protective levels of ND antibody titer with higher 5.0 levels increased from 2.1% in 2001 to 8.8% in 2005.

Efficacy of ELISA for measurement of protective newcastle disease antibody level in broilers (육계의 뉴켓슬병 방어역가 측정에 있어서 ELISA 검사법의 효용성)

  • Kim, Jong-Nyeo;Heo, won;Mo, In-Pil
    • Korean Journal of Veterinary Research
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    • v.46 no.3
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    • pp.185-196
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    • 2006
  • Newcastle disease (ND) is a highly contagious disease of poultry that can cause severe economic losses throughout the world. Vaccination has been used for a long time and proved as one of the most effective method to reduce the economic loss due to ND virus infection, The measurement of antibody titer such as hamagglutination-inhibition (Hl) test with sera has been used as a useful method to evaluate the immunity leve of host. However, Hl test is gradually being replaced by the enzyme linked-immunosorbent assay (ELISA), To evaluate the efficacy of ELISA in the chickens vaccinated with different procedure, present study has been performed. After SPF chicks and commercial broilers were vaccinated with different kinds of live vaccines such as V4, VG/GA and/or Bl at various time, the antibody level has been measured using both HI test and ELISA. Challenge test with velogenic viscerotropic NDV was also performed to measure the protective level of antibody. In the SPF chickens, the mean ELISA titer after vaccination and survival rate after challenge was increased and correlated with days post inoculation. More than 80% of chickens with higher than 1,000 ELISA titer after vaccination were survived after challenge with velogenic ND virus and had good correlation between survival rate and antibody titier. In commercial broiler chickens, most of them at market age had low level of ELISA titer regardless of the number of vaccination, and had a low correlation between survival rate and ELISA titer. However, the ELISA titer of remaining birds after challenge was increased. This result indicated that ELISA titer had good response against velogenic NOV infection compared to Hl titer.

Studies on the production of Japanese encephalitis virus vaccine (일본뇌염백신 제조에 관한 연구)

  • 유건희;이용재
    • Korean Journal of Microbiology
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    • v.9 no.4
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    • pp.175-178
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    • 1971
  • Because of the cases of Japanese Encephalitis(J.E.) were reported every year in Korea. We, Dong-A Pharmaceutical Co., Ltd., produced J.E. virus vaccine, with lower price, since 1970 in order to prevent ourselves from being infected by the disease. And inoculated the J.E. virus vaccine for the children with a great success. We are going to report several questions which brought about in producing the J.E. virus vaccine by alcohol precipitation, protamine sulfate treatment method. The results obtained were as folows ; 1) In process treated with 40% alcohol, we used to ethanol made in Germany, but it was too expensive to use it. As the result which we had studied about it, we were satisfied with J.E. virus vaccine which produced with alcohol made in Korea, and then, we treated with accurate specific gravity of 40% ethanol for the precipitation of the virus. And also, we knew that it was the best method to be treated it for 3hrs, $13^{\circ}C$. 2) When we treated with protamine sulfate (0.025mg/ml), we acquired the highest potent titer, and suited into purpose for the nitrogen concentration. 3) The filtration of the purified J.E. virus vaccine, in case of millipore filter paper of large pore size was not suitable for the sterility. Therefore the pore size less than 0.8.$\mu$ (AA filter paper) in millipore filter paper was very suitable. But it seemed to be important subhects that the smaller was the pore size, the lower was the potent titer.

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