• Title/Summary/Keyword: Viral titer

Search Result 92, Processing Time 0.031 seconds

Identification of Retroviral Vectors Producing High Viral Titer

  • Shin, Yong-Jae;Lenardo, Michael J;Park, Tae-Kyu;Lee, Kwang-Ho
    • The Journal of Korean Society of Virology
    • /
    • v.29 no.1
    • /
    • pp.33-38
    • /
    • 1999
  • Retroviral vector provide a highly efficient method for gene transfer into eukaryotic cells. This vector system can be divided into two components; the retroviral vector itself and the retroviral packaging cell line. The key improvement in the design of these two components are, focused on two aspects; the reduction of helper virus production and high titer-virus. We used PA317 for retrovirus packaging cell line, for its high producibility of viral titer. To test the ability of the vectors to generate high titer-virus, we have chosen four different retroviral vectors; LN, LNSX, LNCX and LXSN. To test easily the viral titer, we have made recombinant construction with CD4 and CD8, checked their viral titer and stained their surface expression. LXSN which contain SV40 early promoter in front of neo gene showed best results in viral transient transfection assay, dot blot assay and surface expression. In addition, recombinant containing CD8 generally showed much higher viral titration and surface expression than CD4.

  • PDF

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
    • /
    • v.52 no.6
    • /
    • pp.644-649
    • /
    • 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.

Prevelance of neutralizing antibody related with viral respiratory disease in cattle (송아지 바이러스성 호흡기 질병 중화항체 조사)

  • Youn, Choong-Keun;Lim, Yeon-Soo;Lyoo, Young S.
    • Korean Journal of Veterinary Research
    • /
    • v.50 no.3
    • /
    • pp.205-211
    • /
    • 2010
  • Fifty young calves, about five to six months old purchased from nation-wide were investigated with the prevelance of neutralizing antibody (Ab) of infectious bovine rhinotracheitis virus (IBRV), parainfluenza 3 virus ($PI_{3}V$), bovine viral diarrhea virus (BVDV) and bovine respiratory syncytial virus (BRSV). The positive detection ratio of neutralizing Ab against IBRV was only 3% and two of positive samples showed low antibody titer (below 2). Ab against BRSV showed 48% of positive ratio and among 24 positive samples, antibody titer of 23 samples were below 3. But in the case of BVDV, 68% of samples were positive and 23 samples appeared to possess high antibody titer, above 4 and the antibody titer of five samples were above 8. The highest positive result came from $PI_{3}V$. The positive ratio in the samples investigated in this study was 72%, but the antibody titer of positive samples were generally below 3 (77.8% in positive samples).

Serological Survey of Cattle on Bovine Viral Diarrhea in Young Dong Province (강원 영동지역 우 바이러스성 설사병의 혈청학적 조사)

  • 이종오;한영도;육심용;김연수;장상문;정재영;김동훈
    • Korean Journal of Veterinary Service
    • /
    • v.14 no.2
    • /
    • pp.148-153
    • /
    • 1991
  • To investigate epidemological sitution of bovine viral diarrhea infection, serological survey in cattle being raised in Young Dong province were conducted. Bovine sera collected ramdomly from August 1990 to December 1990 were tested for bovine viral diarrhea virus serum neutralizing antibody titers. The results were as follows 1. BVDV SN antibody levels were considerably varies and positive rate was 58(108 heads out of 186) 2. BVDV SN antibodies to breeds of cattle was various and positive rates showed that diary cattle, beef, native cattle(Korean) were 67.52%, 59.38%, 27.00% respectively followed in that order. 3. In the regional prevalence of BVD SN antibodies in cattle, Alpine(92%) was the highest, Young Dong south(59%) middle(44%), and North 30% followed in that order 4. In the age relatated prevalence of BVD SN antibodies, the younger than 6 month old group was the highest 65.7%, and older than 25 month old group was also at 62.2%. Then, 7 to 12 moth old group and 13 to 24 month old group showed to 58.5%, 52.1% respectively. 5. The geometric mean titer (log2) of 108 cattle serum samples showing positive BVD SN antibodies was 4.3. 6. In the geometric mean titer(log2) according to age, younger than 6 month old group (5.2) was the highest, then 7 to 12 month old group 2.8(SD=1.94 standard deviation) was lowliest.

  • PDF

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
    • /
    • v.31 no.2
    • /
    • pp.81-85
    • /
    • 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.

Establishment of Plasma Working Standards for the Performance and Quality Assurance of NAT Screening Tests for HIV, HCV and HBV (HIV, HCV와 HBV 유전자 분석시약의 성능 및 품질관리용 Plasma Working Standards 제조에 관한 연구)

  • Kim, Myung Han;Cho, Youn Jung;Kwon, So-Yong;Cho, Nam Sun
    • The Korean Journal of Blood Transfusion
    • /
    • v.23 no.2
    • /
    • pp.152-161
    • /
    • 2012
  • Background: Since Jan. 2012, for performance evaluation of viral reagents, analysis of domestic samples has been recommended in order to obtain approval from the KFDA when they are first introduced to Korea. This regulation requires the standard domestic materials driven from locally infected samples. We tried manufacturing the plasma working standards of HBV, HCV, and HIV NAT using a mixed titer of viral loads. Methods: Forty three HBV DNA positive plasmas, 25 HCV RNA positive plasmas, and 26 HIV RNA positive plasmas were evaluated according to viral load and genotype. Several plasma units, which had high-titer viral loads and the common viral genotypes in Korea, were selected as the source materials for each viral standard. To adjust the appropriate concentration based on the detectable range of variable viral reagents, the source plasma was diluted to several concentrations, divided into small vials, and analyzed for quantification. Results: The 13 plasma working standards, which had variable viral loads for the mixed titer performance panel of HIV, HCV, and HBV NAT, were produced. Conclusion: These national standard materials were first produced in order to supply the mixed titer performance panel for the viral NAT reagent of the level IV transfusion related high-risk group in Korea.

Classifying Host Susceptibility Using Porcine Circovirus Type 2 Viral Load and Antibody Titer (돼지 써코바이러스 2형 감염량과 항체가를 이용한 자돈의 저항성군 선발법)

  • Lim, Kyu-Sang;Lee, Eun-A;Lee, Kyung-Tai;Chun, Taehoon;Hong, Ki-Chang;Kim, Jun-Mo
    • Journal of Life Science
    • /
    • v.27 no.3
    • /
    • pp.283-288
    • /
    • 2017
  • Porcine circovirus type 2 (PCV2) is a notorious and ubiquitous virus in the swine industry. The susceptibility of the host to PCV2 infection is considered to be one factor associated with the dynamics of PCV2. The objective of this study was to verify the criteria for host susceptibility to PCV2, using blood parameters of post-weaned pigs naturally infected with the virus. The PCV2 DNA viral load, antibody titer, and leukopenia characteristics were measured in the serum extracted from the pigs at the 10th week. We classified the pigs into high (>5.0), intermediate (3.0 to 5.0), and low (<3.0) groups on the basis of the PCV2 viral load (log copies/ml), or as positive (${\leq}0.50$) and negative (>0.50) groups on the basis of antibody titer (sample-to-negative corrected ratio). Moreover, using these two categorized parameters, we suggested the criteria for classification into the susceptible and resistant groups. Statistical analyses revealed that pigs in the susceptible group had a significantly higher viral load (p<0.001) and negative antibody titer (p<0.001), as well as significantly lower leukocyte counts (p=0.018) and lower amounts of several leukocyte components (p<0.05), than pigs in the resistant group. We concluded that the susceptible group could be considered to have PCV2-induced leukopenia. Therefore, we suggest that the combined classifications of viral loads and anti-PCV2 antibodies can be used to determine PCV2-induced leukopenia in the subclinical PCV2 infection of post-weaned pig populations.

Genetic association of polymorphisms in porcine RGS16 with porcine circovirus viral load in naturally infected Yorkshire pigs

  • Lee, Seung-Hoon;Lim, Kyu-Sang;Hong, Ki-Chang;Kim, Jun-Mo
    • Journal of Animal Science and Technology
    • /
    • v.63 no.6
    • /
    • pp.1223-1231
    • /
    • 2021
  • Regulator of G protein signaling 16 (RGS16) is known to be associated with porcine circovirus type 2 (PCV2). PCV2 associated disease (PCVAD) is a serious problem in the swine industry. The representative symptoms of PCVAD are high viral titer proliferation and decreased average daily gain. In this study, we identified single nucleotide polymorphisms (SNPs) in the RGS16 region, including the upstream region. Of the 22 identified SNPs, rs332913874, rs326071195, and rs318298586 were genotyped in 142 Yorkshire pigs. These SNPs were significantly associated with the PCV2 viral load. Moreover, the haplotype combination was also related to the PCV2 viral load. The haplotype and diplotype analysis also had a significant difference with the PCV2 viral load. Taken together, our results suggest that RGS16 SNPs considerably affect the PCV2 viral load.

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
    • /
    • v.37 no.4
    • /
    • pp.273-280
    • /
    • 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.

Rapid determination of baculovirus titers an antibody-based assay

  • Kwon, M.S.;Dojimal, T.;Park, Enoch-Y.
    • 한국생물공학회:학술대회논문집
    • /
    • 2003.04a
    • /
    • pp.315-319
    • /
    • 2003
  • A novel method is developed to yield virus titers in 10 h, is easy to .perform using 96-well plates, and applicable to both any Autographa californica nucleopolyhyderovirus (AcNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV)-based recombinant baculovirus. This assay uses an antibody to a DNA-binding protein to detect the infected cells via immune-staining. The titer is determined by counting foci produced due to infection of virus under a fluorescent microscopy. The required incubation period was shortened considerably because infected cells expressed viral antigens at the post infection time of 4 h. Therefore, 10 hours were enough to estimate the virus titer including virus infection time, insect cell culture, and estimation of virus titer.

  • PDF