• Korean Journal of Veterinary Research
• /
• v.32 no.1
• /
• pp.57-61
• /
• 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.

• Korean Journal of Veterinary Service
• /
• v.42 no.1
• /
• pp.9-15
• /
• 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.

• Korean Journal of Veterinary Service
• /
• v.15 no.1
• /
• pp.32-45
• /
• 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.

• Korean Journal of Veterinary Service
• /
• v.17 no.1
• /
• pp.19-24
• /
• 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.

• Korean Journal of Veterinary Research
• /
• v.63 no.4
• /
• pp.37.1-37.7
• /
• 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.

• Journal of Life Science
• /
• v.20 no.10
• /
• pp.1427-1432
• /
• 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.

• The Journal of the Korean Society for Microbiology
• /
• v.3 no.1
• /
• pp.43-49
• /
• 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.

• Korean Journal of Veterinary Research
• /
• v.48 no.1
• /
• pp.83-88
• /
• 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.

• Korean Journal of Veterinary Service
• /
• v.24 no.3
• /
• pp.271-278
• /
• 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.

• Korean Journal of Veterinary Research
• /
• v.30 no.3
• /
• pp.297-307
• /
• 1990

To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.