• Title/Summary/Keyword: PCR-ELISA

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Suggestions for a better inspection method according to the occurrence of fowl typhoid in broiler breeders (육용종계 가금티프스 발생에 따른 검사방법 개선 방안)

  • Chu, Keum-Suk;Kim, Kyoung-Taek;Yoon, Eun-Jeong;Kim, Beom-Seok
    • Korean Journal of Veterinary Service
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    • v.37 no.1
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    • pp.45-49
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    • 2014
  • Pullorum disease and fowl typhoid are septicemic avian diseases transmitting through egg by transovarian infections. A series of tests has been performing in breeding flocks of chickens and test plans for proper inspection have been modified by government veterinary institute to control of such diseases. To improve inspection plans, different test methods were compared using fowl typhoid positive samples from a poultry farm located in Jeonbuk state in 2012. Based on first inspection, 11 samples among total 200 samples were positive by rapid slide agglutination (RSA) test and 7 samples among RSA positive samples were finally diagnosed as Salmonella Gallinarum infection by ELISA, bacterial isolation, PCR, and histopathologic examination. In the second inspection, 20 samples among total 100 samples were positive by RSA test. Among RSA positive ones, 19 samples were positive by ELISA, S. Gallinarm were successfully isolated in 3 samples, and 16 samples were positive by PCR in the cecal tonsils where were not successful for bacterial isolation. Based on histopathologic examination, severe inflammation in the 13 cecal tonsils and infiltration of lymphocytes and heterophils in the 11 livers were observed. Therefore, we suggest that bacterial isolation, PCR, and histopathologic examination methods in the third inspection need to be further used in various tissues for correct diagnosis and for final eradication of pullorum disease and fowl typhoid in breeding flocks of chickens.

Effective Application of Diagnostics for Bovine Leukemia Virus in Dairy Cattle (젖소에서 소 백혈병 진단법의 효과적 활용)

  • Youn, Choong-Keun;Jung, Ho-Kyoung;Sunwoo, Sun-Young;Lyoo, Young-S.
    • Journal of Veterinary Clinics
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    • v.27 no.4
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    • pp.402-406
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    • 2010
  • Bovine leukemia virus (BLV) is a delta-retrovirus which causes chronic lymphocytosis in cattle. BLV infections have been divided into two groups such as enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL) according to the clinical symptoms in infected cattle. The conventional detection method of BLV was hematological procedure which is determining lymphocytosis in the suspected animals. Recently several sensitive methods were developed to detect antibody to BLV and nucleic acid of the BLV from infected cattle. In this study we have compared the difference of positive rates between agar gel immunodiffusion (AGID) and enzyme linked immunosorbent assay (ELISA) which are using for BLV antibody detection methods. The positive detection rate of ELISA test was 7.4% greater than the positive rate of AGID. The discrepancy of the positive rate between ELISA and AGID were showed in the group of age over one year old to under three year old group. The result from each test agreed very well in the group of over 5 year old cattles. The serological test is very useful method to select the infected cattle for the eradication or control of the disease in the infected herd. But it has a limit by interference of the maternal antibody from the cow of under 6 month old. This study shows that 16.2% of these ages group showed BLV gene positive by polymerase chain reaction (PCR) method. The result suggests that ELISA test need to be used with PCR to clarify misinterpretation of positive animals by antibody response due to the natural infection from maternally derived antibody in calves of under 6 months old.

Infection status of bovine viral diarrhea virus in Korean native cattle in Jeonbuk State, Korea (전북지역 한우의 소 바이러스성 설사 바이러스 감염 현황 조사)

  • Woo Ri Jung;Mi Seon Kang;Keum-Suk Chu
    • Korean Journal of Veterinary Service
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    • v.47 no.2
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    • pp.73-79
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    • 2024
  • Bovine viral diarrhea virus (BVDV) is distributed in cattle worldwide and causes significant economic losses to the livestock industry. Identification and remove of BVDV persistently infected (PI) cattle is very important to control BVDV infection in cattle herd. The objective of this study is to investigate the infection status of BVDV infection in Korean native cattle (Bos taurus coreanae) farms located in Jeonbuk State. From 2021 to 2022, a total of 1,497 samples were collected from 17 cattle farms and tested for BVDV antigen using a commercial ELISA kit. By the first ELISA testing, 24 cattle from six farms were positive for BVDV antigen, showing the farm-level or cattle-level prevalence of 35.3% or 1.6%, respectively. By the second ELISA testing which carried out with the first ELISA-positive samples after three-weeks, 12 cattle (0.8%) from five farms (29.4%) were positive for BVDV antigen, indicating these cattle were PI cattle. Genotypes of BVDV were determined with 12 BVDV-positive samples using a previously described RT-PCR assay and the results showed that 3 (25.0%) and 9 (75.0%) were confirmed to be type 1 and type 2, respectively. These results will be helpful to establish the effective control strategy for BVDV in cattle farms in Jeonbuk State.

The anti-allergic effect of SESHINGO(SSG) (세신고가 알레르기성 비염에 미치는 효과)

  • Kim, Min-Ae;Kim, Mi-Bo;Ko, Woo-Shin;Yoon, Hwa-Jung
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.21 no.3
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    • pp.69-81
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    • 2008
  • Objective : The purpose of this study was find out the anti-allergic effects of SESHINGO on the allergic rhinitis. Methods : Cell viability was evaluated by the MTT assay. Cells were treated with the indicated concentration of SSG. $TNF-\alpha$ concentration was measured from cell supernatants using ELISA method. IL-4 concentration was measured from cell supernatants using ELISA method.$IFN-\gamma$ concentration was measured from cell supernatants using ELISA method. Total RNA was isolated, $TNF-\alpha$ and IL-4 mRNA expression was detected by RT-PCR analysis Total RNA was isolated, COX-1, COX-2 mRNA was analyzed by RT-PCR analysis. Results : 1. $\beta$-hexosaminidase release in RBL-2H3 cells were decresed by SESHINGO attentively. 2. Secreting ration of $TNF-\alpha$was restrained in RBL-2H3 cells by SESHINGO attentively. 3. Secreting ration of IL-4 was restrained in RBL-2H3 cells by SESHINGO attentively 4. Revelation of $TNF-\alpha$ mRNA was decresed in RBL-2H3 cells as concetration. 5. Revelation of.IL-4 mRNA was decresed in RBL-2H3 cells as concetration. 6. Revelation of COX-2 mRNA was decresed in RBL-2H3 cells. Conclusion : According to above results, to evaluate the effect of SSG on the anti-allergic effects.

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Comparison of PRRSV and antibody detection in oral fluid and serum samples from different age categories of PRRSV endemic farms (PRRS 양성농장의 사육단계별 구강액과 혈액을 이용한 PRRSV와 항체 검출 비교)

  • Kim, Jung-Hee;Son, Jae Guk;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.43 no.3
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    • pp.173-179
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    • 2020
  • The objective of this study was to evaluate the usefulness of detection of PRRSV and PRRSV-specific antibodies in oral fluids for monitoring of PRRSV infection in endemic farms. The level of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in five age groups of pigs (6, 9, 12, 16 weeks of age and gilts). The samples (25 serums and 5 oral fluids/per a farm) were collected from 5 different farms endemically infected by PRRSV. Both serum and oral fluid samples were tested for PRRSV by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and for anti-PRRSV antibodies by two commercial PRRSV ELISA kits. ELISA mean s/p ratios (2.98 vs 1.63) and positive rate (84.0% vs 68.8%) of the oral fluid samples showed significantly higher levels but had similar patterns to the seroprofile of the blood samples. The PRRSV positive rate of oral fluid and serum samples was 40.0% and 44.0% respectively. In conclusion, the use of oral fluids for PRRS monitoring in endemic farms is strongly recommended.

Response of fetal rat calvarial cells on mineral trioxide aggregate after IL-$1{\beta}$ stimulation (IL-$1{\beta}$ 처리 백서 두개관 세포의 mineral trioxide aggregate에 대한 반응)

  • Lee, Sool-Heon;Park, Ji-Il;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.39 no.3
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    • pp.359-365
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    • 2009
  • Purpose: The purpose of this study was to investigate the ability of Mineral trioxide aggregate(MTA) to support osteoclastic differentiation from fetal rat calvarial cell. Methods: In this study, response of IL-6, RANKL, and OPG in fetal rat calvarial cells stimulated with IL-$1{\beta}$ on MTA was evaluated by ELISA and RT-PCR. Results: The results were as follows; there was no significant difference between glass and MTA at 5days. In ELISA analysis, Glass group and MTA group showed similar IL-6 expression, Glass+IL-$1{\beta}$ group and MTA+IL-$1{\beta}$ group showed similar IL-6 expression. In RT-PCR analysis, Glass group and MTA group showed similar IL-6, RANKL, OPG mRNA expression, MTA+IL-$1{\beta}$ group and Glass+IL-$1{\beta}$ group showed 3 fold increase of IL-6 and RNAKL mRNA expression when compared with MTA group. All groups showed similar OPG mRNA expression. Conclusions: MTA does not suppress cell proliferation and increase the proinflammatory cytokine that induce osteoclastogenesis. Thus, MTA is biocompatible material that could be used in various clinical conditions.

Analysis of Genetically-Modified Soybean and Soybean Sprout by Enzyme-Linked Immunosorbent Assay (ELISA) (효소면역측정법을 이용한 유전자재조합 콩과 콩나물의 분석)

  • Kwak, Bo-Yeon;Ko, Seung-Hee;Shin, Won-Sun;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.556-560
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    • 2003
  • It was determined whether the sandwich ELISA using specific anti-CP4 EPSPS polyclonal and monoclonal antibodies, developed in the previous study, could be applied to detect GM soybean or not. The soybeans (47 imported and 20 domestic soybeans) were analyzed by a sandwich ELISA. The results of imported soybeans were divided into two groups which were high contents $(39.1{\pm}13.5\;{\mu}g/g,\;n=33)$ and low contents of CP4 EPSPS $(2.6{\pm}1.2\;{\mu}g/g,\;n=14)$. The ratio of GM in imported soybeans was about 70.2%. One the other hand, the contents of CP4 EPSPS in domestic soybeans was very low $(0.9{\pm}0.5\;{\mu}g/g,\;n=20)$ which determined to be non-GM soybeans. In case of soybean sprouts, the contents of CP4 EPSPS in soybean sprouts were different between GM and non-GM soybean sprout. The CP4 EPSPS in cotyledon of GM soybeans sprout was higher than that in root hair. The contents of CP4 EPSPS in soybeans sprout of domestic soybeans were very low. Thus, it was possible to determine that the soybeans sprout was made of GM or non-GM soybeans. Also, PCR experiment showed that the sandwich ELISA was accurate to distinguish the soybeans to be GM or non-GM. These results showed the sandwich ELISA could determine the soybeans were GM or non-GM, rapidly and simply.

Iridovirus infection of cultured juvenile flounder (Paralichthys olivaceus) in nursery (종묘장 넙치 치어에서의 Iridovirus 감염)

  • Kim, Tae-Jung;Jang, Eun-Jin;Kim, Jong-Su;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.46 no.1
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    • pp.21-25
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    • 2006
  • Iridovirus is an icosahedral cytoplasmic double-stranded DNA virus with a genome size of 170-200kb. Outbreaks of fish iridovirus infection are characterized by their wide geographic distribution and broad host spectrum, especially in water temperatures of $25-27^{\circ}C$ Recently, the causative agent of high mortalities in flounder (Paralichthys olivaceus) was identified as fish iridovirus in Korea. Iridoviral infection repeatedly occurs in the same area for long periods, suggesting the possibility of viral infection in nursery. To examine this, the existence of iridovirus in juvenile flounders was detected by PCR using virus-specific primers. Antibodies induced against this virus were also examined using ELISA. As a result, juvenile fish in nursery were found to be previously infected with iridovirus, suggesting that proper prevention systems are required.

Application of Oral Fluid Sample to Monitor Porcine circovirus-2 Infection in Pig Farms (구강액을 이용한 양돈장의 Porcine circovirus-2 감염에 대한 모니터링)

  • Kim, Won-Il
    • Journal of Veterinary Clinics
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    • v.27 no.6
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    • pp.704-712
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    • 2010
  • Porcine circovirus-2 (PCV2) has been implicated in many clinical diseases/syndromes that are now referred to as PCV-associated diseases (PCVAD). Due to significant economic losses caused by PCVAD, many swine operations have launched extensive monitoring programs for PCV2. Traditional serum sampling is, however, rather expensive and laborious, hampering effective large scale pathogen surveillance. A field-based longitudinal study was conducted to assess the utility of pen-based oral fluid sample as an alternative to serum for herd PCV2 testing. Six pens (25 pigs/pen) at each of 3 different sites were used in the study. One oral fluid and 5 random serum samples per pen were collected at 3, 5, 8, 12, and 16 weeks of age, and the sera were pooled by pen for testing. All samples were tested for PCV2 by real-time PCR and for antibodies by indirect fluorescent antibody test (for both anti-PCV2 IgG and IgA) and 3 ELISA assays (blocking ELISA, indirect ELISA, and IgG/IgM sandwich ELISA). PCV2 DNA was detected in oral fluid samples sporadically until 8 weeks and in all pens at 16 weeks. PCV2-specific IgG was detected in oral fluid samples at 3 weeks and persisted until 5 to 8 weeks in all sites. Anti-PCV2 IgG and IgA were detectable in oral fluid samples collected at 16 weeks from all of the pens at 1 site. The detection of PCV2 and anti-PCV2 antibody in oral fluid samples correlated positively with results on pooled sera, suggesting that oral fluids can be a cost-effective alternative to serum for herd monitoring of PCV2 infection.

Monitoring of Nervous Necrosis Virus (NNV) in the Broodstock of Seven Band Grouper Hyporthodus septemfasciatus (신경괴사증바이러스(Nervous Necrosis Virus, NNV) 모니터링을 통한 무감염 능성어(Hyporthodus septemfasciatus)친어의 선발)

  • Kim, Si-Woo;Kim, Wi-Sik;Seo, Han-Gill;Kim, Kyong Min;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.527-533
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    • 2017
  • We investigated the infection of nervous necrosis virus (NNV) in seven band grouper Hyporthodus septemfasciatus broodstocks, which have been reared in aquaculture farms in South Korea during 2012-2014. To investigate the prevalence of NNV within the broodstock, egg, sperm, and blood were sampled in the spawning season. The egg and sperm samples were subjected to a nested reverse transcription (RT) polymerase chain reaction (PCR) assay to detect NNV and were inoculated on SSN-1 cells to culture the virus. Blood samples were used to detect antibodies against NNV using enzyme linked immunosorbent assay (ELSIA). Positive values from ELISA were found in 39 of 162 samples (24%) in 2012, and 13 of 28 samples (46%) in 2014. Additionally, 4 of 34 broodstocks (11%) investigated in 2013-2014 were determined to be carriers from the nested RT-PCR and in vitro cultivation. The broodstocks in which antibodies against NNV were detected by ELISA, or in which NNV was detected by the nested RT-PCR assay, posed a risk of vertical transmission of NNV. Therefore, it is necessary to select virus-free broodstocks in seed production to reduce the possibility of the vertical transmission of NNV.