• Title/Summary/Keyword: Serum neutralization test

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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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Development of monoclonal antibody capture ELISA for the detection of antibodies against transmissible gastroenteritis virus

  • Oh, Yeonsu;Tark, Dongseob
    • Korean Journal of Veterinary Service
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    • v.42 no.1
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    • pp.9-15
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    • 2019
  • Transmissible gastroenteritis (TGE) is a disease confined to pigs of all ages, and can be a significant cause of economic loss in breeding herds, primarily because of the very high piglet mortality. The causative agent is a coronavirus, an enveloped positive strand RNA virus and closely related but non-enteropathogenic porcine respiratory coronavirus (PRCV). Although the TGEV has declined with its innocent relative, PRCV, further genome changes could not be excluded. Therefore, the herd-level immunity against this virus is important for the prevention of disease and should be carefully monitored. The aim of this study is to develop monoclonal antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) which can rapidly and accurately determine a large numbers of serum samples for surveillance purpose, and to compare the ELISA with a TGEV-specific serum neutralization test. The MAC-ELISA was sufficiently achieved, and the comparison with the virus-specific serum neutralization assays for 713 sera from pig farms showed a high correlation ($r^2=0.812$, P<0.001). The specificity and sensitivity of MAC-ELISA for the serum neutralization test 91.9% and 91.6%, respectively, which means that the antibody detected by the MAC-ELISA could be said to be protective antibodies. In conclusion, the developed MAC-ELISA would be very helpful in evaluating protective antibodies against TGEV.

Detection of Antibody to Infectious Laryngotracheitis Virus by Enzyme Linked Immunosorbent Assay (효소면역법에 의한 닭 전염성 후두기관염 바이러스 항체 측정에 관한 연구)

  • 임숙경;위성하;최정옥;고홍남
    • Korean Journal of Veterinary Service
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    • v.15 no.1
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    • pp.32-45
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    • 1992
  • In order to establish and enzyme-linked immunosorbent assay to ILTV, field virus strain of ILTV was propagated in chorioallantoic membrane of the embryonated eggs. purified and used as antigen. The antisera selected from the field samples and immunized chickens based on serum neutralization test were used as the standard positive and negative sera in all tests. It was found that optimal antigen concentration was $2{\mu}g$ of protein per well and a 1 : 100 dilution of standard serum showed low background optical density with negative serum and high P/N values of positive sera. A 1 : 500 dilution of the rabbit anti-chicken IgG peroxidase conjugate produced a high P/N values and thirty minutes was chosen as suitable time to read the optical density of the enzyme substrate reaction and optical density was consistent during the 16 hours after stopper was treated. When coated antigen was kept on microplate for varying time up to 16 hours at $4^{\circ}C$ or $37^{\circ}C,$ no significant difference was observed between the treatment. The coated antigen could be kept without change of antigenicity for at least one month at $-70^{\circ}C,\; -20^{\circ}C,\; 4^{\circ}C$ and room temperature. When blocking buffer contanining bovine serum albumin was mixed directly with conjugate and serum at 10% level induced higher P/N values compared to blocking antigen coated microplate with the same blocking buffer. The coefficience of correlation between ELISA and SN test was 0.577. When antibody response of chickens, vaccinated with ILTV, was examined by ELISA and SN test, antibody rising and decay pattern between the two test was similar until 11 weeks of age. However 12 weeks onward antibody titer checked on by SN test was slightly lower than that tested by ELISA.

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Survey on Actually Infected Condition of Aujeszky′s Disease to the Consigned Pigs in Seoul from 1990 to 1993 (90~93년도 서울 지역에 출하된 돼지의 Aujeszky′s병 감염 실태 조사)

  • 최준식;육동현;김성삼;문현칠
    • Korean Journal of Veterinary Service
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    • v.17 no.1
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    • pp.19-24
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    • 1994
  • The porcine Aujeszky's disease was surveyed by Enzyme Immunodiffusion method and serologic neutralization test to the slaughtered pigs at slaughtehouse only in Seoul from March, 1990 to October, 1993. After detecting the positive by enzyme immunodiffusion method primary, we decided finally the positive by serologic neutralization test secondary. Results obtained through the experiments were summarized as followed; 1. The positive of Aujeszky's disease in March, 1990 was 2 of 1,000 sera. 2 positive were decided as the consigned pigs in Kalsan, Hongsung, Chungnaa 2. The positive of Aujezsky's disease in May, 1991 was decided 1 serum at Pogok, Yongin, Kyeonggi. 3. All of the positive detected by Enzyme immunodiffusion Method in 1993 were decided finally in the negative sera. 4. The positive sera detected in 1992 were decided 32 sera at Gyeonggi, 6 at Chungnam, and 1 at Gangwon. Especially, the positive sera percentage detected by Kit Latex Aujeszky Test appeared 78.04% at Gyonggi and by enzyme immunodiffusion Method appeared 11.11% at Chungnam and Gangwon.

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Detection of immunity in sheep following anti-rabies vaccination

  • Hasanthi Rathnadiwakara;Mangala Gunatilake;Florence Cliquet;Marine Wasniewski;Mayuri Thammitiyagodage;Ramani Karunakaran;Jean-Christophe Thibault;Mohamed Ijas
    • Clinical and Experimental Vaccine Research
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    • v.12 no.2
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    • pp.97-106
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    • 2023
  • Purpose: Rabies is a fatal but preventable disease with proper pre-exposure anti-rabies vaccination (ARV). Dogs, as household pets and strays, are the reservoir and vector of the disease, and dog bites have been associated with human rabies cases in Sri Lanka over the past few years. However, other susceptible species having frequent contact with humans may be a source of infection. One such species is sheep and immunity following ARV has never been tested in sheep reared in Sri Lanka. Materials and Methods: We have tested serum samples from sheep reared in the Animal Centre, Medical Research Institute of Sri Lanka for the presence of anti-rabies antibodies following ARV. Sheep serum samples were tested with Bio-Pro Rabies enzyme-linked immunosorbent assay (ELISA) antibody kits used for the first time in Sri Lanka and our results were verified by a seroneutralization method on cells (fluorescent antibody virus neutralization, FAVN test) currently recommended by World Organization for Animal Health and World Health Organization. Results: Sheep received annual ARV and maintained high neutralizing antibody titers in their serum. No maternal antibodies were detected in lamb around 6 months of age. Agreement between the ELISA and FAVN test, i.e., coefficient concordance was 83.87%. Conclusion: Annual vaccination in sheep has an effect on maintaining adequate protection against rabies by measurements of anti-rabies antibody response. Lambs need to be vaccinated earlier than 6 months of age to achieve protective levels of neutralizing antibodies in their serum. Introducing this ELISA in Sri Lanka will be a good opportunity to determine the level of anti-rabies antibodies in animal serum samples.

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.

Production of the Antiserum against Recombinant Envelop Protein, rVP466 for the Neutralization of White Spot Syndrome Virus (WSSV) (흰반점바이러스(WSSV)의 중화를 위한 재조합단백질 rVP466의 항혈청 생산)

  • Gong, Soo-Jung;Kim, Yeong-Jin;Choi, Mi-Ran;Kim, Sung-Koo
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1427-1432
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    • 2010
  • This study was carried out to evaluate neutralization effects against WSSV using antiserum produced from recombinant envelop protein, rVP466 of WSSV. The VP466 gene of WSSV was cloned into pCold I expression vector and rVP466 was expressed in E. coli RIPL. The antiserum against rVP466 was produced in white rabbits (New Zealand white rabbit). The specific immunoreactivity to the antigen, rVP466, was confirmed by Western blot. The constant amounts of WSSV at $1{\times}10^4$ diluted stocks were mixed with various antiserum concentrations and then injected to the muscle of shrimp, Penaeus chinensis, for the neutralization challenge. The shrimps challenged with WSSV as a positive control and those with the mixture of WSSV and preimmune serum as a preimmune control showed 100% cumulative mortality at 17 days post challenge and 83% at 25 days post challenge, respectively. The shrimps challenged with 3 different mixtures of WSSV and rVP466 antiserum at ratios of 1:0.01, 1:0.1 and 1:1 showed 73%, 53% and 46% cumulative mortalities at 25 days post challenge, respectively. These results indicated that WSSV could be neutralized by the rVP466 antiserum. These results suggest that envelop protein VP466 is involved in the initial step of WSSV infection in shrimp.

Multiplication and Antibody Formation of Japanese Encephalitis Virus in Snakes - 1. Antibody responses to the virus and serum

  • Lee, Ho-Wang;Kee, Ryong-Sook
    • The Journal of the Korean Society for Microbiology
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    • v.3 no.1
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    • pp.43-49
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    • 1968
  • Japanese encephalitis(JE) shows its explosive epidemicity in the temperate zone of Asia but little is known on the overwintering mechanism. One of the hypotheses on the overwintering mechanism is that the virus overwinters in the hibernating animals. There has been no report on the proliferation of JE virus(JEV) or antibody formation in the snakes. The purpose of this experiment is to explore the mutual relationship between JEV and snake and to clarify whether JEV proliferates and induce antibody formation in snakes. Three species of non-poisonous common snakes were employed. Precipitation test was carried out after injecting calf serum and, HI and neutralization tests were done by injecting JEV into the snakes. The gamma globulin fraction of pre- and post-injection serum were compared by paper chromatography. According to the results, precipitating antibody reaction to calf serum could be observed only at $4^{\circ}C$. It was failed to demonstrate HI antibody formation but neutralizing antibody could be detected in one of the 9 snakes. Although antibody could not be detected in test-tube, tile result of paper chromatography shows the remarkable increase of gamma globulin fraction after the injection. Above results are strongly indicating the antibody formation in the snakes.

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Prevalence and genotypes of pestivirus in Korean goats

  • Yang, Dong-Kun;Kweon, Chang-Hee;Kim, Byoung-Han;Choi, Cheong-Up;Kang, Mun-Il;Hyun, Bang-Hun;Hwang, In-Jin;Lee, Cheong-San;Cho, Kyoung-Oh
    • Korean Journal of Veterinary Research
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    • v.48 no.1
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    • pp.83-88
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    • 2008
  • In total, 1,142 serum samples were collected from 223 goat flocks rising in five different regions of Korea. These samples were screened for the presence of border disease virus (BDV) antibodies using an enzyme linked immunosorbent assay. Of the 1,142 samples, we found 47 bovine viral diarrhea virus (BVDV) positive cases (4.1%). These positive serum samples were also examined further by using the virus neutralization test against BDV. In addition, samples were tested for both BVDV and classical swine fever virus (CSFV). All of the samples that were seropositive for BDV also demonstrated positive antibody titers against BVDV and CSFV. Due to their common antigenicity, we also determined further the prevalence and carried out virus neutralization test against three pestiviruses: 314 of the goat samples were screened using reverse transcription polymerase chain reaction with primer pairs specific to common pestivirus genome regions. Overall, 1.6% (5/314) of the samples tested was positive for pestivirus. Based on the nucleotide sequence data and the phylogenetic analysis, three isolates were characterized as BVDV type 1 and two isolates as BVDV type 2. However, none of the isolates could be classified as BDV. These results indicate that BVDV-1 and BVDV-2 are the pestivirus strains circulating among Korean goat populations.

Sero-epidemiology of transmissible gastroenteritis of pigs in Gyeongbuk province (경북지방 돼지의 전염성 위장염에 대한 혈청학적 역학조사)

  • 조광현;박최규;김영환;박인화;김성국;박노찬;정종식
    • Korean Journal of Veterinary Service
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    • v.24 no.3
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    • pp.271-278
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    • 2001
  • Swine sera collected from January to December 2000 were tested for the survey on transmissible gastroenteritis(TGE) infection in Gyeongbuk province. Serum neutralization(SN) test was peformed in finishing pigs of 50 pigs industry without clinical signs of TGE and sows, gilts and growing pigs of four pigs industry with TGE. As the results of SN test in fifty industry, antibody titers of 90.6%(474/523 samples) showed below 4. Antibody titers of them showed highly in two pigs industry of western region and three pigs industry of southern region of Gyeongbuk province. The results indicated that TGE had been occurred before in five pigs industry. Most of antibody titers showed of highly in four pigs industry having had TGE. There were no significant differences of the antibody titers of TGE according to age when the survey was made. The above results indicated that the pigs of Gyeongbuk province were not almost exposed in porcine respiratory coronavirus(PRCV) by present.

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