• Korean Journal of Veterinary Research
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• v.42 no.1
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• pp.73-80
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• 2002

An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.

• Journal of Veterinary Clinics
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• v.23 no.4
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• pp.416-420
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• 2006

Compare to testing sera individually, pooled-serum testing has considered as a cost-effective method, particularly on a large population-based seroprevalence studies. This study was to determine the relationship between individual sera and pooled sera titers for detection of avian infectious bronchitis virus (IBV) and to evaluate suitability of pooled sera by comparing prevalences estimated from both samples. A total of 5,000 individual samples were collected from 500 flocks in Chungcheong, Gyunsgi, and Kangwon provinces between January 2005 and February 2006. Ten samples were randomly selected from each flock. Five-hundred pooled sera were prepared by mixing equal amount of each 10 individual serum from the original samples. IBV antibody titers were measured by hemagglutination inhibition (HI) test. The least squares regression analysis was performed to construct equation between pooled and mean individual titers. To determine whether the flock is infected 4 arbitrary criteria were used: detection of at least 1 chicken with HI titer ${\ge}$ 9 (criterion 1), detection of at least 2 samples with HI titer ${\ge}$9 (criterion 2), detection of at least 1 sample with HI titer ${\ge}$ 10 (criterion 3), and filially detection of at least 1 sample with HI titer ${\ge}$ 11 (criterion 4). The receiver operating characteristic (ROC) curve was used to examine the cut-off points of pooled titers showing optimal diagnostic accuracy. The area under the curve (AUC), sensitivities (Se), specificities (Sp), and positive (PPV) and negative (NPV) predictive values were calculated. The regression equation between pooled titers (pool) and mean individual titers (mean) was: $pool= 1.2498+0.8952{\times}mean$, with coefficient of determination of 87% (p< 0.0001). The optimal cut-off points of pooled titers were titer 8 for criterion 1 (AUC=0.975, Se=0.883, Sp=0.959, PPV=0.985, NPV=0.728), titer 8 for criterion 2 (AUC=0.969, Se=0.954, Sp=0.855, PPV=0.926, NPV=0.907), titer 9 for criterion 3 (AUC=0.970, Se=0.836, Sp=0.967, PPV=0.978, NPV=0.772), and titer 9 for criterion 4 (AUC= 0.946, Se=0.928, Sp=0.843, PPV=0.857, NPV=0.921). The difference of 'prevalence estimated by individual and pooled sample showed a minimum of 2% for criteria 2 and a maximum of 9.1:% for criteria 3. These results indicate that the use of pooled sera in HI test for screening IBV infection in laying hen flocks is considered as a cost-effective method of testing large numbers of samples with high diagnostic accuracy.

• Korean Journal of Veterinary Research
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• v.45 no.4
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• pp.569-574
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• 2005

Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.329-338
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• 2007

Samples collected from 15 broiler farms(47 flocks, 920 1-day-old chicks) during March to December, 2006, To survey serum antibody titers of NDV, IBDV and MG/MS, the antibodies of ND viruses were detected by HI test and ELISA, against antibodies of IBD viruses and MG/MS by ELISA. The antibody titers of NDV showed 6.4, HI and 6,968, ELISA, respectively. The rate to below protective antibody levels(${\ge}5$, HI and ${\ge}1,000$, ELISA) were 8%, HI, 5%, ELISA, specially, Baeksemi were 22%, HI, 14%, ELISA. The rate of positive by ELISA showed 99%(914/920). The ELISA titer of IBDV showed mean titer 3,890. The rate of positive were 93% (857/920), specially, Baeksemi were 84%. The ELISA titers of MG/MS showed mean titer 5,666. The rate of positive were 78% (715/920) and 100%, Abor-Acre, 97%, Baeksemi, respectively. The antibodies not detected from 18%, ELISA titers was varied from 500 to 20,000. At antimicrobial susceptibility of E coli, Staphylococcus spp and Salmonella spp isolated from 1-day-old chicks, E coli were susceptible to AmC, AM, NOR, SXT, ENR, CIP, Staphylococcus spp were susceptible to AmC, SXT, AM, ENR and Salmonella spp were susceptible to AM, AmC, SXT and P.

• 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.

• Korean Journal of Veterinary Service
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• v.28 no.2
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• pp.121-146
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• 2005

Newcastle disease (ND), infectious bronchitis (IB), low pathogenic avian Influenza (LPAI) and fowl typhoid (FT) have been known as egg drop laying diseases because of the serious layer damage from mass zone layer. In this study, such egg drop laying diseases were investigated. To access this study, we peformed to evaluate antibody titers in serum and isolated bacteria and virus from organs and feces on May, July and September in 2003. The distribution of ND from January to May, IB and LPAI from October to February of the next year, and FT from March to September were inspected by the question survey in 21 farms. ND revealed to be positive rates of 490 to 474 $(96.7\%)$ in May, 510 to 506 $(99.2\%)$ in July and 510 to 510 $(100\%)$ in September with hemagglutination inhibition (HI) test. The mean antibody titers were 10.2, 9.9 and 10.2, respectively. With regard to IB, 484 out of 490 samples $(98.7\%)$ in May, 508 of 510 $(99.6\%)$ in July and 509 of 510 $(99.8\%)$ in September showed positive results and the mean antibody titers were gradually increased with 8.2, 9.0 and 9.4, respectively. According to HI test of LPAI, the positive results were shown in 442 of 480 $(92.1\%)$, 394 of 494 $(79.8\%)$ and 402 of 483 $(83.2\%)$ in May, July and September, respectively The mean antibody titers were decreased with 4.6, 4.3 and 4.0. The distribution of LPAI also elicited the positive rates of 480 to 475 $(99.0\%)$ in May, 494 to 485$(98.2\%)$ in July, 483 to 472 $(97.7\%)$ in September as determined by ELISA and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. Compared ELISA results with HI test of LPAI the positive results were 480 to 422 $(92.1\%),\;475(99.0\%),\;494\;to\;394 (79.8\%),\;485 (98.2\%)\;and\;483\;to\;402(83.2\%),\;472(97.7\%)$. Therefore, the positive rate determined by ELISA was higher than that of HI test with 6.9, 18.4 and $14.5\%$, respectively. When performed RT-PCR for ND using organ and feces samples, the pathotypes were detected $5(15.6\%)\;in\;May,\; 2(5.3\%) in\;July,\;2(7.1\%)$ in September but there is no samples showing positive band for LPAI. In attempt to isolate Salmonella gallinarum, bacteria were obtained from 4 cases (12.5%) in May, 9 (23.6%) in July, 5 (17.8%) in September. Thus the highest rate for isolation revealed to be shown in July When evaluated the antimicrobial susceptibility to 18 isolated strains of 5. gallinarum, bacteria were sensitive to trimethoprim/sulfamethox$(61.1\%),\;kanamycin\;(55.5\%),\;ampicillin\;(55.5\%)$ and amoxacillin/clavulanic acid $(55.5\%)$, cephalothin $(50.0\%)$, but resistant to penicillin $(88.9\%)$, streptomycin $(88.9\%)$, erythromycin $(83_4\%)$ and tetracycline $(61.1\%)$. According to HI test of ND and LPAI using captured 164 wild Korean tree sparrows (Passer nontanus), the positive rates were $47.6\%\;and\;57.3\%$, and the mean HI titers were 5.32 and 4.02, respectively. 71 $(43.2\%)\;and\;58(35.3\%)$ in captured sparrows also showed more than 4 titers for HI test to ND and LPAI, respectively However, the attempt for isolation of viruses failed in all samples.

• Biomedical Science Letters
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• v.8 no.4
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• pp.269-273
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• 2002

Dengue fever (DF) is an acute febrile illness caused by dengue viruses in the family Flaviviridae, genus Flavivirus. DF has so far posed any problem in Korea, however it has been recently believed to be associated with oversea's traveler infected with dengue virus. Antibody titers of sera from DF patients against dengue virus were measured by indirect immunofluorescence assay (IFA) and plaque reduction neutralization test (PRNT), including the haematologic test. Three of patients with DF showed highly fluorescent and neutralizing antibody titers by IFA and PRNT assay. Two of them showed higher, remarkably. Meanwhile, one of them was tested and resulted in severe tirombocytopenia, elevated serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) activities as well as mild leucopenia, increased monocytes and basophils and depressed lymphocytes in haematological differential count.

• Journal of Veterinary Clinics
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• v.18 no.4
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• pp.368-373
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• 2001

To study the immunomodulatory effect of aquapuncture with canine parvovirus killed vaccine, the vaccine was inoculated into the dogs twice with on 2-week interval. The 6 dogs in the experimental group were inoculated through the Jiao-Chao acupoint, and 5 dogs in the control group were done subcutaneously. The antibody titer was determined by the hemagglutination inhibition test. The HI titers of the experimental group showed significantly higher on days 21 and 28 than those of the control group. The biochemical test on serum total protein, protein fractions and the A/G ration showed a slightly increased in $\gamma$-globulin on days 21 and 28. The hematological findings on total leukocytes and differential counts showed no significance. It was thought that the aquapuncture of the canine parvovirus killed vaccine through the Jiao-Cho acupoint may stimulate the antibody production.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.49-51
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• 1985

A serological survey for antibodies to Leptospira (L.) interrogans in dairy and Korean native cattle in Gyeongbuk area was performed using 6 different living antigens, which were L. icterohaemorrhagiae(RGA), L. canicola(Hond Utrecht IV), L. autumnalis(Akiyami A), L. australis(Ballico), L. pomona(Pomona) and L. hebdomadis(Hebdomadis), by microscopic agglutination test. In the microscopic agglutination test, greater than partial agglutination at a serum dilution of 1:300 or over was recorded as positive. In the dairy cattle(Holstein), four(4.7%) of 86 sera from 13 dairy farms were positive for L. canicola and L. pomona antibodies. In the Korean native cattle, four(3.2%) of 124 sera from the slaughter house in Taegu city were positive for L. canicola, L. icterohaemorrhagiae and L. pomona antibodies. Among the positive sera, L, canicola was dominated in this experiment.

• Korean Journal of Veterinary Research
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• v.2 no.1
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• pp.51-54
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• 1962

Chickens of Leghorn breed were bursectomized at 3 weeks of age and inoculated intranasally at 4 weeks of age with the $B_1$ strain of NDV. There was no statistically significant difference(P>0.1) in the serum titers (HI test) between the bursectomized chickens and the control chickens.