• Title/Summary/Keyword: viral antibodies

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Development of a Blocking ELISA for Measuring Rabies Virus-specific Antibodies in Animals

  • Yang, Dong-Kun;Kim, Ha-Hyun;Ryu, Jieun;Gee, Mi-ryun;Cho, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.269-276
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    • 2018
  • Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

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
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    • v.14 no.2
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    • pp.148-153
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    • 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.

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Detection of antibodies against infectious Borna disease virus -a comparison of three serological methods- (보르나병 바이러스 항체검출을 위한 연구 -세 가지 혈청진단법의 비교-)

  • Lee, Du-sik
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.57-61
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    • 1992
  • To determin the accuracy of serological methods in detecting Borna-disease(BD) viral antibodies, 273 experimentally infected rabbit sera were compared by using indirect immunofluorescence antibody test(IFA), serum neutralization test(SN) and enzyme-linked immunosorbent assay(ELISA). One hundred twenty-three serum samples had BD viral antibodies detected by IFA. CELISA antibodies to BD virus were also present in the same one hundred twenty-three serum samples. However, neutralization test antibodies to BD virus were present in 27 of the in rabbit serum samples. Neutralization test was sensitive in comparison with KFA and CELISA. In comparison with IFA, CELISA was both sensitive and specific in detecting BD viral antibodies. These results extend observations made with laboratory animals to the diagnosis of naturally infected animals.

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Incidence of canine viral diseases and prevalence of virus neutralization antibodies of canine distemper virus, adenovirus type 2, parvovirus, and parainfluenza virus type 5 in Korean dogs

  • Dong-Kun Yang;Ha-Hyun Kim;Hye Jeong Lee;Young-Ju Cheong;Lee-Sang Hyeon;Minuk Kim;Bang-Hun Hyun
    • Korean Journal of Veterinary Research
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    • v.64 no.1
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    • pp.3.1-3.8
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    • 2024
  • Canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus (CPV), and canine parainfluenza virus 5 (CPIV-5) are the major viral pathogens in dogs. Despite the availability of vaccines for dogs against these 4 viral pathogens, investigations of antibodies against these pathogens have rarely been reported in South Korea. In this study, we investigated the recent incidence of viral diseases in dogs and conducted sero-surveillance for CDV, CAV-2, CPV, and CPIV-5 in Korean dogs. The most frequently diagnosed canine viral disease in Korean dog samples from 2000 to 2022 was CPV infection, which accounted for 48.7% (464/953) of the cases. A total of 400 dog serum samples collected between 2019 and 2022 were screened for the presence of virus-neutralizing antibodies against CDV, CAV-2, CPV, and CPIV-5. The overall seropositivity rates for CDV, CAV-2, CPV, and CPIV-5 were 83.8%, 77.8%, 99.3%, and 82.0%, respectively. The protection rate against CPV was the highest (98.3%) and that against CAV-2 was the lowest (44.8%) in dog sera. Male and female dogs showed no significant differences in seropositivity rates. CDV and CPIV-5 seropositivity increased with age in dogs, and the highest incidence and seropositivity rates of CPV indicated that Korean dogs have been continuously exposed to wild CPV, and that CPV is a pathogen that urgently requires attention among canine viral diseases.

Prevalence of antibodies against bovine viral infectious diseases in farmed deer and wild water deer in Jeonbuk province (사육사슴 및 야생고라니의 소 바이러스성 전염병에 대한 혈청학적 연구)

  • Jo, Young-Suk;Chu, Keum-Suk;Lee, Jeong-Won;Camer, Gerry A;Chekarova, Irina;Seol, Min-Suk;Park, Hyun-Jong;Kim, Bum-Seok;Lim, Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.32 no.2
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    • pp.111-117
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    • 2009
  • Farmed deer could be susceptible carrier to bovine viral infectious disease. But unfortunately, there has not been an overall study over this subject in Korea so far. Therefore, a study was conducted to see serum antibodies to bovine leukosis, food and mouth disease, bovine viral diarrhea and infectious bovine rhinotracheitis in deer using the sera of farmed deer. As a result, two deer in a farms showed positive in bovine leukosis antibodies, using ELISA. For wild water deer, no antibodies were found for those diseases. As a result, it can be assumed that deer were relatively low rate of exposure to highly contagious disease such as viral bovine infectious disease in Korea. As this study was conducted over limited in number of subject and regions, continued study should be carried out in order to prevent and control the interspecies transmission in the future.

Monoclonal antibodies against structural proteins of bovine viral diarrhea virus (소 설사병 바이러스 구조단백에 대한 단크론항체 성상에 대한 연구)

  • Kweon, Chang-hee;Zee, Yuan Chun;Woo, Hee-jong
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.83-90
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    • 1992
  • Monoclonal antibodies against structural proteins of bovine viral diarrhea virus(BVDV) were derived by classical hybridoma techniques. These antibodies were characterized by serum neutralization, immunoblotting and immunoprecipitation. The neutralizing monoclonal antibody reacted with the 56kd to 54kd(M.W.) viral protein in western blotting and immunoprecipitation analysis. Although there was no neutralizing activity, another monoclanal antibody reacted with the 45kd protein by immunoprecipitation and with both the 45kd and 36kd proteins in immunoblotting analysis. respectively. Densitometer scanning of purified BVDV and the immunopreipitation of whole virus particles with neutralizing monoclonal antibody revealed the presence of more than twelve viral polypeptides. Although no possible precursor form of protein was identified with the neutralizing monoclonal antibody. the presence of intact virion was detected in the infected cell supernatant immediatelty after pulse labeling, indicating rapid translational processing as well as packaging of the virus. The partial peptide mapping of 45kd and 36kd proteins with Staphylococcus aureus V 8 protease showed that these two proteins are related.

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Detection of Specific Antibodies Against Viral Hemorrhagic Septicemia Virus in Infected Olive Flounder Paralichthys olivaceus Using Enzyme-Linked Immunosorbent Assay (Enzyme-linked immunosorbent assay를 이용한 바이러스성 출혈성 패혈증 바이러스 감염 넙치(Paralichthys olivaceus)의 특이 항체반응 검사)

  • Hwang, Jee Youn;Jang, Jin Hyeon;Kim, Dong Jun;Kwon, Mun Gyeong;Seo, Jung Soo;Hwang, Seong Don;Son, Maeng-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.547-552
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    • 2017
  • The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.

Improvement of indirect enzyme-linked immunosorbent assay for detection of Japanese encephalitis virus antibodies in swine sera

  • Yang, Dong-Kun;Kim, Ha-Hyun;Jo, Hyun-Ye;Lee, Seung Heon;Jang, Sang-Ho;Lee, Sang-Oh;Choi, Sung-Suk;Cho, In-Soo
    • Korean Journal of Veterinary Research
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    • v.57 no.1
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    • pp.31-36
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    • 2017
  • Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

Evaluation of hemagglutination inhibition test for canine respiratory coronavirus antibodies and seroprevalence in Korean dogs

  • Lee-Sang Hyeon;Dong-Kun Yang;Yu-Ri ,Park;Hye Jeong Lee;Ha-Hyun Kim;Bang-Hun Hyun
    • Korean Journal of Veterinary Research
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    • v.63 no.4
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    • pp.37.1-37.7
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    • 2023
  • Canine respiratory coronavirus (CRCoV) is a significant pathogen that causes respiratory diseases in dogs, collectively known as a canine infectious respiratory disease. The virus is highly contagious and exhibits high seroprevalence worldwide. Currently, bovine coronavirus (BCoV) enzyme-linked immunosorbent assay (ELISA) kits are used to detect CRCoV antibodies. However, BCoV-ELISA kits cannot differentiate between infections caused by BCoV and those caused by CRCoV. In this study, we evaluated the hemagglutination inhibition (HI) test for CRCoV by comparing it with the virus neutralization (VN) test. Subsequently, we evaluated the seroprevalence of CRCoV in 383 dog serum samples collected from South Korea utilizing the HI test. The HI test for CRCoV showed a strong correlation with the VN test (R = 0.83, p < 0.001). The analysis of seroprevalence revealed that 52.2% (95% confidence interval [CI], 47.2%-57.1%) of the Korean dog serum samples were positive. The seroprevalence exhibited varied with age, with a positivity rate of 43.9% in dogs under 1 year of age and 66.7% in dogs aged 3 to 5 years (odds ratio, 2.54; 95% CI, 1.43-4.59). In conclusion, the HI test to monitor CRCoV antibody proved to be closely related to the VN test. Furthermore, over half of the dogs in Korea tested positive for CRCoV antibodies. These findings contribute to a better understanding of the sero-epidemiology of CRCoV.

Prevalence of autoantibodies that bind to kidney tissues in cats and association risk with antibodies to feline viral rhinotracheitis, calicivirus, and panleukopenia

  • Songaksorn, Nisakorn;Petsophonsakul, Wilaiwan;Pringproa, Kidsadagon;Lampang, Kannika Na;Sthitmatee, Nattawooti;Srifawattana, Nuttawan;Piyarungsri, Kakanang;Thongkorn, Kriangkrai
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.38.1-38.17
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    • 2021
  • Background: The feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP) vaccine, prepared from viruses grown in the Crandell-Rees feline kidney cell line, can induce antibodies to cross-react with feline kidney tissues. Objectives: This study surveyed the prevalence of autoantibodies to feline kidney tissues and their association with the frequency of FVRCP vaccination. Methods: Serum samples and kidneys were collected from 156 live and 26 cadaveric cats. Antibodies that bind to kidney tissues and antibodies to the FVRCP antigen were determined by enzyme-linked immunosorbent assay (ELISA), and kidney-bound antibody patterns were investigated by examining immunofluorescence. Proteins recognized by antibodies were identified by Western blot analysis. Results: The prevalences of autoantibodies that bind to kidney tissues in cats were 41% and 13% by ELISA and immunofluorescence, respectively. Kidney-bound antibodies were observed at interstitial cells, apical border, and cytoplasm of proximal and distal tubules; the antibodies were bound to proteins with molecular weights of 40, 47, 38, and 20 kDa. There was no direct link between vaccination and anti-kidney antibodies, but positive antibodies to kidney tissues were significantly associated with the anti-FVRCP antibody. The odds ratio or association in finding the autoantibody in cats with the antibody to FVRCP was 2.8 times higher than that in cats without the antibody to FVRCP. Conclusions: These preliminary results demonstrate an association between anti-FVRCP and anti-cat kidney tissues. However, an increase in the risk of inducing kidney-bound antibodies by repeat vaccinations could not be shown directly. It will be interesting to expand the sample size and follow-up on whether these autoantibodies can lead to kidney function impairment.