• 한국임상수의학회지
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• 제17권1호
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• pp.13-20
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• 2000

Recently cases of canine distemper have occurred in Korea despite vaccination was carried out nationwidely. This study was performed to establish rapid diagnosis of canine distemper by RT-PCR, nested PCR, and serological test. A total of 30 dogs, which were suspected canine distemper clinically, was examined. RT-PCR and nested PCR were specific for the amplification of CDV H gene and sensitive to detect 7 TCID50 of Onderstepoort strain. By RT-PCR, H gene was detected in 6(20%) of 30 peripheral bloods from dogs. And H gene was detected in 10(33.3%) of 30 samples by nested PCR. H gene was detected from 1 brain of 6 years-old Beagle dog and 1 lung of 2 months-old Shihtzu dog, in which peripheral blood H gene was not detected. Serum neutralizing antibody titer against Onderstepoort strain ranged from 4 to 1,024 in 30 patients. No correlation was observed between the results of nested PCR and titiers of neutralizing antibody.

• 대한수의학회지
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• 제25권1호
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• pp.69-75
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• 1985

Hog cholera serum neutralizing antibody of piglets and sows were titrated by means of END SN method. The piglets of a variety of ages, precolostrally immunized with LOM living HC vaccine were subjected to the test. The sows were vaccinated with lapinized living HC vaccine after 25 days from the parturition. Throughout the studies the following results were obtained and summarised. 1. Hog cholera antibody titers of inbred sows immunized with lapinized living HC vaccine after 25 days from parturition were high except Hampshire group(Table 2). 2. Sows, different stage of the pregnancy or the day of parturition, and of 3 way crossed, that were immunized with lapinized living HC vaccine have shown no significant difference on HC antibody titer(Table 2, 3). 3. HC antibody titers of piglets, immunized with a single dose of LOM HC vaccine before feeding colostrum, were high in case of the younger group(1 week) compare to the older(7 week) (Table 4). 4. The piglets that were booster immunized with LOM HC vaccine at the age of 7 weeks have shown an inconsistent antibody titers(Table 5).

• 대한수의학회지
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• 제43권3호
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• pp.449-456
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• 2003

The VP2 gene of aquatic birnavirus, Korean isolate (GC-1) was cloned and expressed using the baculovirus expression system. The VP2 gene and VP2 partial gene, which contained a neutralizing epitope, were constructed for recombinant transfer vectors, for baculovirus expression. The expressed recombinant proteins were confirmed by indirect immuno fluorescence antibody (IFA), SDS-PAGE and Western blot. The level of expression was checked at regular time using IFA and Western blot. To measure the neutralizing activity of recombinant proteins against GC-1 strain, the antisera against recombinant proteins were produced by using guinea pigs. The result showed that the antisera neutralized the GC-1 strain. However, the neutralizing titer was higher in antisera against the VP2 gene expressed recombinant protein than that of VP2 partial gene recombinant protein.

• 대한수의학회지
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• 제48권1호
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• pp.53-60
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• 2008

We have investigated efficiency of a recombinant subunit Pasteurella multocida toxin (PMT) that was mixed with a vaccine consisted of inactivated whole cells of Bordetella bronchiseptica, P. multocida (types A and D). For verification of the efficacy of the vaccine, all experimental pigs (suckling piglets, sow and gilts) in the three farms were vaccinated. Antibody titers against B. bronchiseptica and P. multocida type A of the vaccinated pigs by microplate agglutination were significantly higher than those of the control pigs (p < 0.05). Similar patterns were observed in the analysis of anti- PMT neutralizing antibody by serum neutralizing method using Vero cell (p < 0.05). Anti- P. multocida type D antibody titer of the vaccinated sows and gilts by ELISA showed significant differences with those of the non-vaccinated pigs (p < 0.05). Although antibody titers increased, it was unable to find out the difference in the clinical signs between the vaccinated and non-vaccinated pigs. However, the increase in body weight of the vaccinated piglets was observed in comparison with the non-vaccinated piglets on a farm. At slaughtering of the pigs, pathological lesions in the turbinate bones of the vaccinated pigs were significantly lower than those of the non-vaccinated pigs (p < 0.001). These results suggested that efficacy of the vaccine in pigs demonstrated to protect against atrophic rhinitis in Korea.

• Journal of Veterinary Science
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• 제22권4호
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• pp.56.1-56.10
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• 2021

Background: Fluorescent antibody virus neutralization (FAVN) test is a standard assay for quantifying rabies virus-neutralizing antibody (VNA) in serum. However, a safer rabies virus (RABV) should be used in the FAVN assay. There is a need for a new method that is economical and time-saving by eliminating the immunostaining step. Objectives: We aimed to improve the traditional FAVN method by rescuing and characterizing a new recombinant RABV expressing green fluorescent protein (GFP). Methods: A new recombinant RABV expressing GFP designated as ERAGS-GFP was rescued using a reverse genetic system. Immuno-fluorescence assay, peroxidase-linked assay, electron microscopy and reverse transcription polymerase chain reaction were performed to confirm the recombinant ERAGS-GFP virus as a RABV expressing the GFP gene. The safety of ERAGS-GFP was evaluated in 4-week-old mice. The rabies VNA titers were measured and compared with conventional FAVN and FAVN-GFP tests using VERO cells. Results: The virus propagated in VERO cells was confirmed as RABV expressing GFP. The ERAGS-GFP showed the highest titer (10^8.0 TCID₅₀/mL) in VERO cells at 5 days post-inoculation, and GFP expression persisted until passage 30. The body weight of 4-week-old mice inoculated intracranially with ERAGS-GFP continued to increase and the survival rate was 100%. In 62 dog sera, the FAVN-GFP result was significantly correlated with that of conventional FAVN (r = 0.95). Conclusions: We constructed ERAGS-GFP, which could replace the challenge virus standard-11 strain used in FAVN test.

• 대한수의학회지
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• 제49권3호
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• pp.201-205
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• 2009

In this study, we evaluated the stability and potency of live attenuated rinderpest vaccines of lapinized-avianized tissue culture strain origin, which had been produced annually from 2005 to 2008. When immune responses to the vaccines were evaluated using two Holstein calves weighing 100~150 kg, neutralizing antibody titer of 1 : 16 was induced at 21 days post vaccination. When calves were also inoculated with vaccines lots that had been stored for 39 months at ${4^{\circ}C}$, same level of antibody titer was observed. Using the virus titer test, we found that all batches of the vaccine that had been kept for 3, 10, 15, 22, 27, 34, 39, and 45 months showed no significant loss of titers, and fulfilled the requirement necessary ($\geq$ 3 $logTCID_50$) to be used as the national rinderpest vaccine reserve in Korea. In this study, we demonstrated that stability and potency of the rinderpest vaccines were maintained over three years when kept at ${4^{\circ}C}$ storage. This indicates that it maybe feasible to extend the expiration period of this vaccine from one year to three years.

• Clinical and Experimental Vaccine Research
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• 제13권3호
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• pp.253-258
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• 2024

Purpose: The immunogenicity of vaccines containing the canine adenovirus (CAdV) type 2 (CAdV-2) variant has not yet been reported. We prepared a novel inactivated CAdV-2 variant vaccine using the CAV2232-41 strain, and evaluated its safety and immunogenicity in raccoon dogs. Materials and Methods: The growth kinetics of CAV2232-41 were determined using Madin-Darby Canine Kidney (MDCK) cells. The nucleotide sequences of CAV2232 and CAV2232-41 were determined by next-generation sequencing. To generate the CAdV-2 variant vaccine, CAV2232-41 propagated in the MDCK cells was inactivated with 0.1% formaldehyde. Two vaccines were prepared by blending inactivated CAV2232-41 with Cabopol and Rehydragel adjuvants. Safety and immunogenicity of the CAV2232C and CAV2232R vaccines were evaluated in guinea pigs. Safety and immunogenicity of the CAV2232C vaccine were also evaluated in raccoon dogs. The virus neutralizing antibody (VNA) titer against CAV2232-41 was measured in sera collected from immunized guinea pigs and raccoon dogs. Results: CAV2232-41 showed the highest viral titer on days 4-6 post-inoculation and had a deletion in the E3 gene, which was confirmed as a CAdV-2 variant. Guinea pigs inoculated with CAV2232C showed slightly higher VNA titers than those inoculated with CAV2232R 2 weeks after booster vaccination. Raccoon dogs immunized with the CAV2232C vaccine developed high mean VNA titers, while non-vaccinated raccoon dogs were antibody-negative. Conclusion: The CAV2232C vaccine is safe and induces a protective VNA titer in raccoon dogs.

• BMB Reports
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• 제37권5호
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• pp.556-564
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• 2004

A recombinant Fab monoclonal antibody (Fab) C37, previously obtained by phage display and biopanning of a random antibody fragment library against Burkholderia pseudomallei protease, was expressed in different strains of Escherichia coli. E. coli strain HB2151 was deemed a more suitable host for Fab expression than other E. coli strains when grown in media supplemented with 0.2% glycerol. The expressed Fab fragment was purified by affinity chromatography on a Protein G-Sepharose column, and the specificity of the recombinant Fab C37 towards B. pseudomallei protease was proven by Western blotting, enzyme-linked immunosorbent assay (ELISA) and by proteolytic activity neutralization. In addition, polyclonal antibodies against B. pseudomallei protease were produced in rabbits immunized with the protease. These were isolated from high titer serum by affinity chromatography on recombinant-Protein A-Sepharose. Purified polyclonal antibody specificity towards B. pseudomallei protease was proven by Western blotting and ELISA.

• 한국동물위생학회지
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• 제32권3호
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• pp.201-207
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• 2009

Porcine transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV) and rotaviruses are considered as the most important causative agents of diarrhea in piglets. The study established 3 method vaccination programs to prevent PEDV. A (LL)group inoculated twice vaccinations on 2-weeks-interval during the late term of pregnant sows with PEDV live vaccine. The B (LKK) group was applied that one time single PEDV live vaccine at the pre-mate followed by the TGEV PEDV combined inactivated vaccine (twice vaccination on 2-weeks interval at the third-trimester). C (KK) group was applied to sow which inoculated twice vaccination on 2-weeks-interval during the late term of pregnant sows with by the TGEV, PEDV combined inactivated vaccine. As the result of SN test on sows in the pig farm before vaccination, antibody titers was showed 9/45 (20.0%). By comparison with the serum neutralizing antibody titers against PEDV of the vaccination programs after PEDV of the vaccination, A group and B group vaccination method was higher than those of C group in sows. In the piglets up to 2 weeks of age, A group was showed antibody titers of 17/22 (81.8%) that showed 2-128, and B group was showed antibody titers of 30/37 (81.1%) that showed 2-512, and C group was showed antibody titers of 14/28 (50.0%) that showed 2-32. On the other hand, PEDV antibody titers were tested for the survey. As the results of SN test, Aujeszky's disease survey in 54 pig farms from november 2005 to august 2006, antibody titers of 47/286 (16.4%) showed above 2. Five breeding farms were antibody titers of 38/77 (49.4%), Wanggung zone farms antibody titers of 59/85 (69.4%). In pigs farms vaccinated the first of twice PEDV live vaccine, and after 6 month, the second of twice TGEV PEDV combined inactivated vaccine (LLKK, 256-1024 titer) method was higher than those of vaccinated twice the early term of pregnant, and twice the late term of pregnant sows of PEDV live vaccine (LLLL, 32 titer).

• 대한미생물학회지
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• 제22권1호
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• pp.1-8
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• 1987

Hantaan virus(HV) 76-118 strain was inoculated into suckling ICR mice by intra-nasal route with an inoculum of $10LD_{50}$. Mortality was 65% at the 3rd week after inoculation, but declined to 35% at the 4th week. Infectivity was determined by the measuring immuno-fluorescent antibody in sera. The peak of infectivity was 80% at the 4'th week after inoculation. Viremia was reached peak level of $1.7{\times}10^4\;PFU/ml$ by day 10. Immunofluorescent antibody and neutralizing antibody appeared by 2 weeks and 15-17 days respectively, but achieved similar titer by 35 days. By using a monoclonal antibody to HV 76-118, viral antigens were initially detected in inguinal and axillary lymph node by 2 days. Viral antigens in bone marrow and lung were delayed much more than in those of lymph node. These were similar with those of intra-peritoneal and intra-muscular route. Immune complex against IgG, IgM and C3 appeared by 16 days, 14 days, and 18 days respectively. The pattern of immunofluorescence in the basement membrane of glomeruli was diffuse membranous. Spotted pattern was also observed in the tissue stained with anti-mouse C3 antibody. By 20 days, control tissue was also shown immune complex in the glomeruli.