• Title/Summary/Keyword: serum HI test

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Comparison of ELISA and HI titers in broiler chicks vaccinated with infectious bronchitis virus and Newcastle disease virus (전염성기관지염 및 뉴캣슬병 백신을 접종한육계에서 ELISA 및 HI 항체가 비교)

  • 고원석;이정원;곽길한;권정택;송희종
    • Korean Journal of Veterinary Service
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    • v.24 no.1
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    • pp.21-29
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    • 2001
  • To compare of serum antibody titers using ELISA and HI, serum samples were collected from 100 breeders and their progeny 550 broilers. The breeders and broilers were vaccinated with infectious bronchitis(IB)- and Newcastle disease(ND)-viruses according to general vaccination program. The antibodies in serum samples against IB and ND viruses were detected by enzyme-linked immunosorbent assay(ELISA) using commercial ELISA kit and hemagglutination inhibition(HI) test. Geometric mean titer(GMT) of ELISA and In titers were monitored from 1-day-old to 35-day-old broilers and compared to those of breeder chickens. The antibody titers of breeders vaccinated with ]B virus showed 47,800, ELISA and 7.2, HI, respectively. Progeny chicks, 1-day-old, vaccinated with IBV showed high antibody titers than those of breed chickens. Those chicks were maintained protective antibody levels until 11-day-old. From 14-day-old, the antibody level decreased below protective levels. In ND, breeders serum antibody titers ELISA and Eiu were 30,200 GMT and 8.7 HI titer, respectively. On 1-day-old chicks, antibody levels was decreased to half in ELISA(16,270) compared with those of breeders, but In titers was 7.4. Progeny broilers, protective antibody level was maintained until 14- day-old by ELISA, but at 11-day-old by HI titers. After then, ND antibody titer was continuously decreased underdefense level. These result indicated that the ELISA method be more sensitive than HI titration to detect serum antibody level for IBV and NDV.

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Studies on porcine parvovirus inactivated vaccine and titration of serum neutralizing antibody (돼지 parvovirus불활화(不活化) vaccine 및 중화항체가측정(中和抗體價測定)에 관한 연구(硏究))

  • Kwon, Hyock-jin;Yeh, Jae-gil;Lee, Chang-koo
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.355-359
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    • 1988
  • A porcine parvovirus inactivated vaccine was prepared and inoculated to 7 piglets and also 8 guinea-pigs, and their serum antibodies were titrated. Twenty-two field serum samples of unvaccinated sows were also tested by SN and HI methods. It was observed that SN test was superior over HI test. Therefore, it is suggested that the SN test could well be used in the detection of serum antibody for PPV in vaccinated pigs.

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Harmonization of Hemolysis Index in Clinical Chemistry Laboratory and Its Application as a Result Verification Tool (화학검사 결과의 검증을 위한 용혈 지수 일치화 도구 개발)

  • Pyo, Sang Shin;Nam, Hyun Su;Cha, Young Jong;Lee, Seungkwan;Lee, Hae Kyung
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.350-358
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    • 2017
  • The hemolysis index (HI) is semi-quantitative marker for hemolysis. Because the characteristics of the HI vary from one commercial platform to another, no standardization or harmonization of the HI is currently available. Specimens (N=40) randomly selected from clinical patients were artificially hemolyzed in vitro. The serum of the specimens was then diluted with a 20 mg/dL difference between 0~300 mg/dL based on serum hemoglobin measured using the XE-2100 hematology automation equipment (Sysmex, Japan). Diluted serum was measured using the Hitachi-7600 biochemical automation equipment (Hitachi, Japan) to differentiate between HI and serum hemoglobin. The data showed linearity between HI and serum hemoglobin and that HI 1 contained approximately 20 mg/dL of serum hemoglobin. To determine the blood rejection threshold, the HI was divided into three groups: HI 0~1, HI 4~6, HI 9~15. After another batch of clinical specimens (N=40) was measured using a Hitachi-7600 (Hitachi, Japan), each specimen was moved forward and backward with the piston of the syringe to induce an artificial in vitro hemolysis, then measured again with a Hitachi-7600 (Hitachi, Japan). The percentage difference between the three groups was analyzed by ANOVA or the Kruskal-Wallis test. In the post-test, there were significant differences between the HI 0~1 and the HI 5~6: Glucose, creatinine, total protein, AST, direct bilirubin, uric acid, phosphorus, triglyceride, LDH, CPK, Magnesium, and potassium levels. Because many clinical tests differed significantly, the threshold for hemolysis could be appropriate for HI 5 (serum hemoglobin 100 mg/dL).

Expression of the VP2 protein of feline panleukopenia virus in insect cells and use thereof in a hemagglutination inhibition assay

  • Yang, Dong-Kun;Park, Yeseul;Park, Yu-Ri;Yoo, Jae Young;An, Sungjun;Park, Jungwon;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.19.1-19.7
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    • 2021
  • Feline panleukopenia virus (FPV) causes leukopenia and severe hemorrhagic diarrhea, killing 50% of naturally infected cats. Although intact FPV can serve as an antigen in the hemagglutination inhibition (HI) test, an accidental laboratory-mediated infection is concern. A non-infectious diagnostic reagent is required for the HI test. Here, we expressed the viral protein 2 (VP2) gene of the FPV strain currently prevalent in South Korea in a baculovirus expression system; VP2 protein was identified by an indirect immunofluorescence assay, electron microscopy (EM), Western blotting (WB), and a hemagglutination assay (HA). EM showed that the recombinant VP2 protein self-assembled to form virus-like particles. WB revealed that the recombinant VP2 was 65 kDa in size. The HA activity of the recombinant VP2 protein was very high at 1:215. A total of 143 cat serum samples were tested using FPV (HI-FPV test) and the recombinant VP2 protein (HI-VP2 test) as HI antigens. The sensitivity, specificity, and accuracy of the HI-VP2 test were 99.3%, 88.9%, and 99.3%, respectively, compared to the HI-FPV test. The HI-VP2 and HI-FPV results correlated significantly (r = 0.978). Thus, recombinant VP2 can substitute for intact FPV as the serological diagnostic reagent of the HI test for FPV.

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.

Serologlcal survey of infections agents in domesticated boars

  • Cho, Kwang-Hyun;Park, In-Hwa;Kim, Young-Hoan;Kim, Soon-Tae;Kim, Sung-Kook;Park, No-Chan;Son, Jae-Kwon;Jyeong, Jong-Sik
    • Korean Journal of Veterinary Service
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    • v.24 no.4
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    • pp.359-367
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    • 2001
  • A serological survey was performed to establish basic data for the prevalence of antibodies to some major diseases of domesticated boar serum samples from January to December 2000. Sera collected in breeding farms in Gyeongbuk province were tested for Aujeszky's disease virus(ADV), Porcine reproductive and respiratory syndrome virus(PRRSV), Porcine parvovirus(PPV), Japanese encephalitis virus (JEV), Bordetella bronchiseptica(B bronchiseptica), Mycoplasma ; APP), Toxoplasma, and Brucella. There was no antibody to ADV in domesticated boars serum samples detected by Anti-ADV-gpI assay kit. Sero-positive samples to PRRS by IFA were 0.9%(3/330) The HI titers to PPV ranged variously from less than 10 to over 1,280. Two hundred ninety-four out of 330 tested sera showed HI titer of less than 10. In HI test to JEV, 90.3% of the sera (298/330) were below 10. The majority of the serum samples had low prevalence of the antibody B bronchiseptica. ELISA titers to M hyopneumoniae ranged variously from $\leq$ 10 to $\geq$ 1,280. Antibody titers to A pleuropneumoniae type 2(APP2) and type 5(APP5) were 58.2% and 52.7%, respectively, and the tested samples showing ELISA antibody titers of less than 20. There was no significant geographical difference between APP2 and APP5 in this study. In the antibody test of Toxoplasma, 11.5%(38/330) were positive and samples were all negative in sera test of Brucella.

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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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Enzyme Linked Immunosorbent Assay for the Detection of Antibody to Avian Infectious Bronchitis Virus (효소면역법에 의한 닭 전염성 기관지염 바이러스 항체검사)

  • 송현제;최정옥
    • Korean Journal of Poultry Science
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    • v.18 no.3
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    • pp.183-196
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    • 1991
  • In order to establish ELISA method to detect antibody against IBV various factors involved were examined. Antigen was prepared from Massachusetts type IBV which is known to be one of serotypes distributed most widely. The virus was grown in embryonated SPF chicken eggs. Allantoic fluid harvested was processed to ultracentrifugation and sucrose density gradient centrifugation to produce a purified antigen The antisera selected from the field samples based on hemagglutination inhibition test were used as the standard positive and negative sera for this study and the results obtained were summarized as follows. 1 , It was found that ELISA test was satisfactory when the purified antigen was coated on the plate in the amount of about 40ng protein per well. In case of the phospholipase treated hemagglutinating antigen it gave satisfactory results when the each well wns coated with 1.2 to 2.5 hemagglutinating unit which was equivalent to 40 to 90ng of protein. 2. There was no significant difference in the ratio of optical density of positive to that of negative serum whether the coated antigen was held for 1 hour at 37$^{\circ}C$ or it was held overnight at 4$^{\circ}C$. The coated antigen could be kept in dried state without change of antigenecity for at least one month of experimental period at 4$^{\circ}C$. 3. There was a big variation in the optical density and P/N values depending on the maker of the plates and on the plate of the same maker. 4. It was found that background optical density was negligible when serum was diluted more than 1:50 and serum dilution of 1:100 appeared to be appropriate as a routine test dilution to screen the antibody. 5. Optical density was fairly constant 15 minutes afterward from the time substrate was treated and during the 4 hours after stopper was treated. 6. There was a low correlation(r=0.42) between ELISA and HI test. However, when 74serum samples were tested for the IBV antibody, 98.7% were found to be positive by both tests in which titers of 2$^{6}$ or more by HI test and P/N values of 1.4 or more by ELISA were considered to be positive, 7 Day-old IBV vaccinated chickens shows a similar antibody decay and rising pattern until 8 weeks of age by the two tests, ELISA and HI.

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Effect of Serum Antibody on Infectious Bronchitis Virus and Its Pathogenicity to SPF Chicks (닭 전염성 기관지염 바이러스에 대한 항체 및 병원성에 관한 연구)

  • Kim Soon-Jae
    • Journal of the korean veterinary medical association
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    • v.25 no.5
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    • pp.282-291
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    • 1989
  • Incidence of infectious bronchitis virus(IBV) infection in vaccinated breeder chickens was investigated by hemagglutination inhibition(HI)test for IBV using Mass 41 antigen. In the breeder chickens with the reduced egg production. chalky deposit. wrinkled

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Effect of Modified Live Virus Vaccine aganist Canine Parvovirus Infection (개 파보바이러스 감염에 대한 Modified Live Virus Vaccine의 효과)

  • 한정희;유기일;권혁무;서강문
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.46-55
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    • 1998
  • This study was performed to verify the effect of modified live virus vaccine against canine parvovirus (CPV) infection. Serum hemagglutination inhibition (Hl) test, histopathological and immunohistochemical techniques and polymerase chain reaction were used. The results were as follows: 1. During the experimental terms after vaccination, serum Hl titer was stable. Geometric mean titer (GMT) after the 1st vaccination was 280. After virulent CPV was challenged, GMT was 1,306. 2. After challenge by virulent CPV, the vaccinated group was not shown clinical signs and gross and histopathological findings. 3. After challenge by virulent CPV, the vaccinated group was not detected viral antigens in the small intestine immunohistochemically. 4. After challenge by violent CPV, the vaccinated group was not shown virus shedding in feces. In conclusions the overall results confirmed that modified live virus vaccine was effective on prevention of canine parvovirus infection.

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