• 대한수의학회지
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• 제46권3호
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• pp.185-196
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• 2006

Newcastle disease (ND) is a highly contagious disease of poultry that can cause severe economic losses throughout the world. Vaccination has been used for a long time and proved as one of the most effective method to reduce the economic loss due to ND virus infection, The measurement of antibody titer such as hamagglutination-inhibition (Hl) test with sera has been used as a useful method to evaluate the immunity leve of host. However, Hl test is gradually being replaced by the enzyme linked-immunosorbent assay (ELISA), To evaluate the efficacy of ELISA in the chickens vaccinated with different procedure, present study has been performed. After SPF chicks and commercial broilers were vaccinated with different kinds of live vaccines such as V4, VG/GA and/or Bl at various time, the antibody level has been measured using both HI test and ELISA. Challenge test with velogenic viscerotropic NDV was also performed to measure the protective level of antibody. In the SPF chickens, the mean ELISA titer after vaccination and survival rate after challenge was increased and correlated with days post inoculation. More than 80% of chickens with higher than 1,000 ELISA titer after vaccination were survived after challenge with velogenic ND virus and had good correlation between survival rate and antibody titier. In commercial broiler chickens, most of them at market age had low level of ELISA titer regardless of the number of vaccination, and had a low correlation between survival rate and ELISA titer. However, the ELISA titer of remaining birds after challenge was increased. This result indicated that ELISA titer had good response against velogenic NOV infection compared to Hl titer.

• 한국동물위생학회지
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• 제26권3호
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• pp.215-219
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• 2003

This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA) titers and sample to positive ratio (S/P ratio) of Newcastle disease(ND) virus. To perform this study, the 372 sera of broiler chicks and 120 sera of layers and breed chicks were collected from slaughter house and farms, respectively. As a result of HI test out of different chicks, the positive percentage of ND antibody titer of broiler, layer and breeder, when a standard positive HI titer were '2', was 84.4%, 100% and 100%, respectively. The positive percentage of ND antibody titer by ELISA was shown 38.4%, 100% and 100% and S/P ratio were also shown 81.5%, 98.2% and 99.2%, respectively. The results of comparative survey with same sera by two experimental methods were as follows; In low HI titer, ELISA titer was not similar to HI titer, but S/P ratio was similar to it. In high HI titer, ELISA titer was not similar to HI titer. Therefore, HI titer was more similar to S/P ratio than ELISA titer.

• 한국환경보건학회지
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• 제23권4호
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• pp.73-81
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• 1997

Diphtheria was the world-spread, respiratory infectious disease in the past. But after developing the diphtheria toxoid, diphtheria vaccine, composed of diphtheria toxoid, is generalized, so prevalence of diphtheria is sharply dropped. But diphtheria is come back in NIS(New Independent States) in 1990s. It is believed that recurrence of diphtheria is mainly due to shortage of protective diphtheria antibody titer in adults. And it is related to 70% prevalence in adults, not in children. Considering this changing environment, this study was designed to research the diphtheria antibody titer of whole population and existence of titer difference by sex. Also it was studied whether ELISA has fitness as determination method of diphtheria antibody titer compared with neutralization method on microcell culture. This study was done to 277 samples of three hospitals in Seoul area, and sera was tested by neutralization method on microcell culture and ELISA method. The results of this study were as follows 1) Of this studied population, 31.05% had an antibody titer below the protective level (<0.01 IU/ml), 40.79% had a titer of a relative degree of protection (0.01-0.09 IU/ml) and 28.16% had a reliable degree of protection ($\geq$0.1 IU/ml). Therefore, 68.95% had a basic protective antibody titer level. 2) 20-40 age group showed the lowest diphtheria antibody titer among the studied population. 3) GMT of diphtheria antibody titer was the highest in the children. But, after child period, GMT was fallen. Standard deviation value was the lowest in older group. 4) Protective level of diphtheria antibody titer of male was slightly higher than female (70.94% vs 66.66%). But this difference was not statistically significant. 5) Diphtheria antibody titer by ELISA method was wholly higher than by neutralization method on microcell culture. Compared the two results by ELISA and neutralization, the regression coefficient was 0.38. And in titer, which was obtained by ELISA method, false-positive results was abundant.

• 대한미생물학회지
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• 제21권2호
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• pp.205-210
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• 1986

Serological investigations for the leptospirosis on hospitalized patients in Choonchun Sungsim Hospital during the periods from August to November 1985 and 841 inhabitants of Kangwondo area including Choonchun, Choonsung, Inje, Chulwon, Hwachun, Gosung, Taibaik, Samchuk and Yangju area were carried out. 1. Among 58 hospitalized patients who were suspected as leptospirosis, 10 patients were detected to have antibody against Leptospira. All of positive sera had the highest antibody titer against serogroup Icterohemorrhagiae and most positive sera were also reactive to serogroup Australis and Canicola. Antibody titer of positive sera detected by microscopic agglutination(MA) test were ranging from 1 : 40 to 1 : 2,560. Antibody titer detected by ELISA method were higher than those detected by MAT(ELISA 1 : 400$\sim$1 : 25,600) and IgM titer of positive sera were generally higher than IgG titer. 2, Of 841 inhabitants in 8 area of Kangwondo, 17 persons (2,02%) possessing antibody against Leptospira were detected by ELISA method, IgG titer in positive sera were generally higher than IgM titer. Persons possessing antibody to Leptospira were distributed in both sex and in various age group, and no significant regional and occupational fluctuations were obserbed.

• 한국동물위생학회지
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• 제21권3호
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• pp.277-284
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• 1998

Serum samples were collected from 100 breeders and their progeny 600 broilers. The breeders and broilers were vaccinated against infectious bronchitis(IB), infectious bursal disease(IBD) and Newcastle disease(ND) viruses according to general vaccination program. The antibodies in serum samples against IB, IBD and ND viruses were detected by ELISA using commercial ELISA kit. Geometric mean titer(GMT) of ELISA was monitored from 1-day-old to 35-day-old broilers and compared to that of breeder chickens. The GMT of ELISA to IB, IBD and ND was declined half level of the day old broiler's antibody titers at about 4, 9 and 4 days of age. The GMT of ELISA to IB, IBD and ND was declined than that of protective antibody titer at about 12, 11, and 15 days of age. Thereafter, the GMT of ELISA to IB, ND were declined and disappeared according to age of broilers. The GMT of ELISA to IBD was declined according to age of broilers, but at 25 days of age increased and 31 days of age increased than that of protective antibody titer. Taken together, these studies led to conclusion that time-course of antibody titers of broilers from vaccinated breeders and that of progeny broliers which vaccinated according to vaccine program. Those are very important data to design vaccine program to breeders and broilers.

• 대한수의학회지
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• 제29권4호
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• pp.503-515
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• 1989

Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay(ELISA) for detection of antibodies to avian infections bronchitis virus(IBV) were standardized. By adopting the optimized conditions an equation calculating ELISA antibody titers from the observations at single serum dilution was formulated. The purified antigen of IBV Mass-41 strain was dispensed into polystyrene microplate wells at a concentration of 300ng per well($100{\mu}l$) and the plates were coated by completey drying at $37^{\circ}C$. Diluted chicken serum and horseradish peroxidase conjugated goat anti-chicken IgG were added in order in $100{\mu}l$ volumes per well and allowed to react for 30 minutes each at room temperature. Just before use and after each reaction the plates were washed three times with distilled water. Finally o-phenylenediamine solution was added as an enzyme substrate. After incubation for another 15 minutes at room temperature absorbances were read at 492nm. Hyperimmune serum against Mass-41 strain was used as internal reference positive(IRP) serum. After repeated titration of IRP and negative serum, a constant titer of IRP was determined. Serum titrations were carried out for various sample sera together with IRP and negative sera and the observed titers of sample sera were corrected by reflecting the ratio between observed and constant titers of IRP serum. These corrected titers of the sample sera were plotted against sample/positive(S/P) OD ratios. All the OD's measured in the serum titrations were also corrected by substracting negative serum OD. The following equation was formulated from the above data; $Log_{10}$ ELISA titer=$5.568({\log}_{10}S/P)+4.161$ Thus it was possible to calculate ELISA titer by measuring absorbance at 1/400 single serum dilution. Titer measured by cross ELISA tests employing Mass-41 strain and three local IBV isolates were similar. These results suggest that the ELISA tests standardized in this study can be used for evaluating not only vaccinal immunity but also for infection status against fields IBV's.

• Parasites, Hosts and Diseases
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• 제30권2호
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• pp.83-90
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• 1992

Toxoplasma증의 혈청학적 진단에 있어서 민감도를 증가시키기 위해 간접 latex 응집반응의 결과와 비교하면서 ELISA를 개발하였으며, 뇌척수액의 검사 시료로서의 가능성을 검토하였다. 아울러 중추신경계 질환환자로부 터기생충질환을 감별하기 위하여 1986년부터 1991년까지 전국 카 병원에서 채취한 혈청과 뇌척수액에 대하여 간접 latex 응집반응(ILA)과 ELISA를 실시하여 Toxoplasma 항체 보유 양상을 비교 검토하였다. 전체 2,016 건의 혈청에 대해 ILA를 실시하여 76건(3.8%)의 양성 (1:32이상의 titer)을 얻었다. 그러나 양성 혈청환자에서 채취한 뇌척수액에서는 낮은 titer의 반응은 있었으나 양성은 나타나지 않았다. 이들 양성 혈청의 양성 혈청 및 음성 혈청에 대하여 ELISA로 항체검사를 실시한바 ILA의 titer가 1 : 32인 군에서 통계적으로 유의한 차이를 나타내는 항체값을 얻었으며, 그 흡반도는 0.40이었다. 뇌척수액에 대한 ELISA로는 ILA의 1 : 64 titer군에서 통계 적으로 유의한 차이가 나타났고 그때의 흡광도 0.27을 양성 판단의 기준으로 사용하였다. ELISA에 의한 항체 검사상 전체 혈청에서 7.0%의 양성을 검출하여 ILA보다 약 2배 정도의 높은 민감도를 보였으며, 뇌척수액에서 는 5.6%의 양성률을 보여 ELISA는 뇌척수액에서의 항체 검출시 유용한 방법이라고 판단하였다. ILA에 비하여 ELISA는 약 2배 정도 높은 양성률을 내었고 양성률은 나이에 마라 40대 이후 급격한 증가를 보였으며, 여성보 다는 남성에서 약 2배 정도 양성률이 높게 나타났다. ELISA에 의한 뇌척수액의 항체 검사에서는 양성률의 성별 차이를 나타내지 않았다. 이상의 결과로 판단할 때, ELISA가 ILA보다 Toxoplasma 항체 검출의 민감도가 높았으며, 뇌척수액은 ELISA의 좋은 검사시료가 되며, 특히 중추신경계 Tocxoplnsma증의 진단에 있어 뇌척수액에 대한 항체 검사에서 ELISA가 유용하다고 판단하였다.

• 한국군사과학기술학회지
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• 제13권3호
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• pp.478-485
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• 2010

The vaccine is biological pretreatment that improves immunity to a particular disease. We can get immunity from producing antibody with injection antigen which has ability to defense against the disease. The ELISA is the most widely used method to measure antibody titer. We have developed and performed validation of ELISA according to the guideline of KFDA and ICH. In this paper, we have verified ELISA method is an excellent method to measure the titer of anti-PA antibody. We have constructed recombinant protective antigen among anthrax toxins and used as antigen of ELISA. In this validation, we have evaluated precision (repeatability, interlaboratory precision), specificity, linearity(range) and LOD, which are validation articles suggested by guideline. Inter-person precision was replaced with inter-laboratory precision. From the results, we have confirmed high precision in all experiments with CV under 20%.

• Parasites, Hosts and Diseases
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• 제25권2호
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• pp.188-194
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• 1987

돼지에 $1.5{\times}10^6$개의 Sarcocystis suicanis sporocyst와 10,000개의 Toxcplasma gondii oocyst(미국 Georgia 대학교 수의과대학 보존주)를 각각 경구감염 시킨후 9주간에 걸쳐 혈중 IgG 항체의 소장과 Sarcocystis와 Tohoplasma 사이의 혈청학적 교차반응성을 IFA 검사와 ELISA법을 이용하여 검사하였다. IFA검사와 ELISA를 위한 Sarcocystis의 전충체 항원과 수용성 항원은 감염돈의 확육을 인공소화액으로 소화시켜 준비하였다. Toxoplasma의 전충체 항원과 수용성 항원은 감염 mouse 복강에서 충체를 분리하여 사용하였다. Sarcocystis와 Togoplasma를 각각 인공감염시킨 돈혈청의 IgG 항체는 IFA검사와 ELISA에서 모두 감염 2주째에 처음으로 검출되었으며 Tcxoplasma에 대한 항체가는 감염 6주째에 최고치에 도달하였다가 그 이후에는 하강하였다. Sarcocystis에 대한 항체가는 감염 9주까지 서서히 상승하였다. Toxoplasma에 감염된 돈혈청은 Sarcocystis 항원에 대하여 IFA검사에서는 1 : 16까지 교차반응이 나타났으며 ELISA에서는 1 : 32까지 나타났다. 대조군의 돈혈청은 모두 1 : 4 이하이었다.

• 한국동물위생학회지
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• 제21권2호
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• pp.133-139
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• 1998

Serum samples collected from 30 breeders and their progeny 30 chicks. The antibodies against infectious bronchitis(IB), infectious bursal disease (IBD) and Newcastle disease(ND) viruses were detected by ELISA using commercial ELISA kit. The breeders were vaccinated against IB, IBD and ND viruses according to general vaccination program. Geometric mean titers(GMT) of ELISA were monitored from 1-day old to 17-day old chicks and compared with breeder chickens. The GMT of ELISA to IB, IBD and ND were declined half level of the breeder antibody titer at 6-, 8- and 7-day old. And, the GMT of ELISA to IB, IBD and ND were declined than that of protective titer at 6-, 1-, and 4-day old. Thereafter, the GMT of ELISA was declined and disappeared according to ages of chicks. Taken together, this study led to conclusion that time-course of maternal antibody titers of chicks from vaccinated breeders, and this is very important data for vaccination to chicks.