• Title/Summary/Keyword: Enzyme immunoassay(EIA)

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Development of an enzyme immunoassay for detection of Escherichia coli O157 in meat (식육중 Escherichia coli O157 검출을 위한 enzyme immunoassay 기법 개발)

  • Jung, Byeong-yeal;Jung, Suk-chan;Cho, Dong-hee;Kim, Jong-yeom;Park, Yong-ho;Shin, Sang-jae;Kim, Sung-guk;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.745-750
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    • 1998
  • A sensitive and rapid enzyme immunoassay(EIA) to detect Escherichia coli O157 in ground beef was developed by using a sandwich type assay with polyclonal antibodies to E coli O157. E coli O157 in ground beef could be detected within 15hr, including incubation for 12hr in enrichment broth and 3hr in immunoassay. The EIA could detect $1.3{\times}10^5$ cells of E coli O157/g of ground beef without enrichment. The lowest limit of detection was 0.23 E coli O157 per g of meat after enrichment. Confirmation was required in the positive specimens in the EIA by culture method even though the negative specimens were not. These results suggested that the immunoassay could be a very efficient method for the screening E coli O157 in meat.

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Studies on enzyme immunoassay for determining progesterone of bovine plasma and its clinical application I. Optimizing double antibody for progesterone in enzyme immunoassay (Enzyme immunoassay(EIA)에 의한 소의 혈중 progesterone 측정과 이의 응용에 관한 연구 I. 이항체(二項體)의 최적조건에 관한 연구)

  • Kang, Chung-boo;Shin, Jong-uk;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.321-325
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    • 1988
  • This experiment was carried out to determine the progesterone concentration of bovine plasma by liquid phase double antibody enzyme immunoassay. The optimum conditions of assay-system, double (first and second) antibody and carrier (normal rabbit serum) were investigated. The optimum dilution rate of first antibody, second antibody and normal rabbit serum was $10{\times}10^3$ to $15{\times}10^3$, 20 and $1{\times}10^3$ times, respectively.

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Prevalence of Anti-HCV among the Health-checkup Adults in Jeonbuk Province (전북 지역 건강 검진자들의 Anti-HCV 양성률 조사)

  • Kim, Yoohyun
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.1
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    • pp.32-37
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    • 2010
  • The author was performed to investigation of current status of prevalence for anti-hepatitis C virus (HCV) among the health-checkup adults in Jeonbuk province. A toal of 1,553 (male 1,046, female 507) serum samples were diagnosed by 3rd generation enzyme immunoassay (EIA) for anti-HCV. Total prevalence of anti-HCV was 0.9%, and prevalence of male and female were 0.8% and 1.2%, respectively. The prevalence of female was higher than male. According to ages group, prevalence of anti-HCV was highest in 60 age group, but it was not found in 20 age group. 14 samples with anti-HCV positive were diagnosed by EIA for hepatitis B virus surface antigen (HBs Ag), by chemiluminescence immunoassay (CLIA) for serum albumin, alanine transaminase (ALT) and asparagine transaminase (AST). Positive for HBs Ag was not found. The mean of serum albumin levels was 4.5 g/dL, and mean of ALT and AST were 34.3 IU and 31.9 IU, respectively. Through this study, I know that the prevalence of anti-HCV among adults in Jeonbuk, and suggest that the positive of anti-HCV persons who have lower serum albumin, normal to mild elevations in serum enzymes are chronic hepatitis.

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Pregnancy Diagnosis in Sows by Using an On-Farm Blood Progesterone Test

  • Wu, L.S.;Guo, I.C.;Lin, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.6
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    • pp.603-608
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    • 1997
  • To improve animal production, a simple and accurate pregnancy diagnosis plays a very important role. Therefore, the purpose of this study was to develop an on-farm blood progesterone enzyme immunoassay (EIA) system for monitoring the early pregnancy in sows. Star tubes coated with mouse monoclonal anti-progesterone antibody were used for this proposed EIA system which was tested in field trials. The results could be obtained within 30 minutes either by spectrophotometry or the naked eye. Heparinized fresh blood samples collected from the ear vein of sows 17-22 days after breeding (day 0) were tested qualitatively to diagnose sows as pregnant or non-pregnant with high ( > 3 ng/ml) or low ($${{\leq_-}}3ng/ml$$) progesterone in the blood. To provided a double check data, plasma progesterone levels were also measured quantitatively by the same EIA system with some modification. Total agreement of diagnosis by the on-farm EIA kit and by farrowing or abortion from 128 tested sows was found to be 92.2% accuracy (93.1% on pregnant diagnosis and 83.3% on non-pregnant diagnosis). It was concluded that the on-farm EIA blood progesterone test is a very useful method for monitoring the early pregnancy status of sows.

Development and validation of a simple, sensitive enzyme immunoassay for quantification of androstenedione in bull plasma

  • Mallick, Smrutirekha;Kumar, BS Bharath;Prakash, BS;Aggrawal, Anjali;Pandita, Sujata
    • Journal of Animal Science and Technology
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    • v.57 no.4
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    • pp.13.1-13.5
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    • 2015
  • As an alternative to radioimmunoassay a simple and highly sensitive enzyme immunoassay (EIA) was developed and validated for androstenedione quantification in plasma of Karan Fries bulls using second antibody coating technique. The wells of the microtitreplate were coated with affinity-purified goat immunoglobulin (antirabbit IgG) that binds the hormone specific antibody. The EIA was performed to analyze androstenedione directly in $40{\mu}l$ of bull plasma. The androstenedione standards ranged from 0.20 to 200 pg/$40{\mu}l$/well and the sensitivity of the assay was 5 pg/ml plasma. Serially diluted bull plasma containing high endogenous androstenedione showed good parallelism with bovine androstenedione standard curve. Intra- and inter-assay coefficients of variation (CV) were found to be 8 and 9%, respectively. Peripheral plasma androstenedione concentrations determined in young and adult bull samples ranged between 104-990 pg/ml and 184-2040 pg/ml, respectively.

Development of an enzyme immunoassay for determination of steroid hormones to improve the reproductive efficiency of domestic animals (가축(家畜)의 번식효율증진(繁殖效率增進)을 위한 steroid hormones 의 효소면역분석법(酵素免疫分析法) 개발(開發))

  • Choi, Han-sun;Kang, Byong-kyu
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.611-615
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    • 1993
  • A rapid, solid-phase microtitre plate enzyme immunoassay(EIA) to determine the concentration of progesterone and testosterone in dairy cow is described. Both steroid hormones were analysed employing antibodies against $11{\alpha}$-hemisuccinate-progesterone bovine serum albumin and 4-androsten-$17{\beta}$-ol-3-one-carboxymethyloxime bovine serum albumin, respectively. as primary antibodies and sheep Ig G as secondary antibody. The conjugated used as labels for progesterone and testosterone was progesterone-$11{\alpha}$-hydroxy-hemisuccinate- horseradish peroxidase and 4 ${\alpha}$-androsten-$17{\beta}$-ol-3-hemisuccinate- horseradish peroxidase, respectively. Detection limit of microtitre plate EIA was 6.7 pg/well for progesteone and 1.0 pg/well for testosterone.

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STUDIES ON THE EARLY PREGNANCY DETERMINATION IN COWS BY USING THE ENZYME-IMMUNOASSY AND RADIO-IMMUNOASSAY IN MILK

  • Lee, J.M.;Kim, H.S.;Jeong, S.G.;Jung, J.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.3
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    • pp.299-302
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    • 1996
  • Milk samples(n = 78) were taken 19d, 20d, 21d, 22d after artificial insemination(AI) for early pregnancy diagnosis by using the Enzyme immunoassay(EIA) kit. The progesterone ($P_4$) concentration in the whole milk was measured on the same day of pregnancy diagnosis. Rectal palpation(RP) was accomplished between 60d and 70d after AI to estimate the ovary condition and pregnancy status. Milk progesterone concentrations measured by Radio-immunoassay(RIA) method, in the pregnant cows at 17d, 19d, 21d after insemination were $17.10{\pm}0.91$, $17.60{\pm}0.46$, and $18.43{\pm}0.79nmol/l$, whereas those in the not-pregnant cows were $6.57{\pm}1.03$, $2.63{\pm}0.29$, and $0.67{\pm}0.08nmol/l$, respectively. When the progesterone concentration was less than 7 nmol/l, the color of the EIA kit was lighter and when the progesterone concentration was ${\geq}16nmol/l$, the color of the EIA kit was darker compared to the standard color. The detection rates of error by judging the color differences were 5.1% and 20.7%, respectively. In the early pregnancy diagnosis by the EIA kit and RIA method, the accuracy rates in the pregnancy of cows were 82% and 87%, and those in not-pregnant cows were 86% and 91%, respectively. For ovarian status estimated by the RIA method and certified by RP, the accuracy rates of the ovarian atrophy, follicular cyst and luteal cyst were 80, 91 and 83% and the progesterone concentrations were 2.51, 2.03, and 26.7 nmol/l, respectively.

Validation of enzyme immunoassay for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae Type b capsular polysaccharide (Haemophilus influenzae type b 피막 다당질 특이 인간 IgG 항체의 정량적 측정을 위한 enzyme immunoassay의 타당성 연구)

  • Kim, Kyung Hyo;Lim, Soo Young
    • Clinical and Experimental Pediatrics
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    • v.50 no.2
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    • pp.143-150
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    • 2007
  • Purpose : This study was conducted to validate enzyme immunoassay (EIA) for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae type b (Hib) capsular polysaccharide. Method : We evaluated specificity, repeatability, intermediate precision, accuracy, lower limit of quantification (LLOQ), and stability to validate standardized EIA for the quantitative measurement of human anti-polyribosylribitol phosphate (PRP) IgG antibodies. Results : The results indicated that this EIA showed specificity to HbO-HA antigen and repeatability and intermediate precision were within acceptance criteria (repeatability: $CV{\leq}15%$, intermediate precision: $CV{\leq}20%$). The EIA-derived results from this laboratory were equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-PRP antibodies in the 28 sera. Spiking recovery result was within acceptance criteria ($100{\pm}20%$). The precision and accuracy of samples in LLOQ were from -14.7 to -4.7% in nominal values, which were within acceptance criteria (precision: $CV{\leq}25%$, accuracy: ${\pm}25%$). Freeze-thaw stability and short term temperature stability were within ${\pm}20%$ of acceptance criteria. Conclusions : The EIA which is performed at the Center for Vaccine Evaluation and Study Ewha Medical Research Institute, is an appropriate serologic assay which can be used for quantitation of anti-PRP IgG antibodies in human sera.

Studies on enzyme immunoassay for determining progesterone of bovine plasma and its clinical application. II. Establishment of enzyme immunoassay for progesterone (Enzyme immunoassay(EIA)에 의한 소의 progesterone 측정과 이의 응용에 관한 연구 II. Progesterone 측정에 대한 효소면역측정방법(酵素免疫測定方法)의 확립)

  • Kang, Chung-boo;Shin, Jong-uk;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.29 no.1
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    • pp.21-25
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    • 1989
  • This experiment was carried out to determine the progesterone concentration of bovine plasma by liquid phase double antibody enzyme immunoassay. The optimum conditions of assay-system, enzyme conjugate and gelatin were invested. The sensitivity, recovery rate and reproducibility by this assay were also analyzed. The results obtained were as follows: 1. The suitable supplementation level of gelatin was 0.2%. As the gelatin level increased to 1%, coefficient variation of interassay was shown to be irregular. 2. The optimum dilution rate of enzyme conjugate was 30 times. Enzyme activity was greatly fluctuated depending on the minor condition of enzyme conjugate technique. Therefore, it was considered to be checked when determined. 3. The sensitivity of the assay was 12 pg/tube. 4. Recovery rate was decreased when the amount of sample was too little or too much, but the recovery rate was high as 97.8% when the amount of sample between 50 and $200{\mu}l$. 5. Mean intra-assay and inter-assay coefficient of variation was 4.5% and 5.9%, respectively. By using liquid phase double antibody enzyme immunoassay, progesterone in plasma can be detected, and also this assay system is applicable to study on physiological function of progesterone and to diagnosis of reproductive disorders.

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Validation of an Enzyme-Immunoassay for Calcitriol in Human Plasma and Evaluation of Its Pharmacokinetics after Single-dose in Korean Volunteers (인체 혈장 중 칼시트리올의 효소면역 분석법 검증 및 단회투여 후 약물동태 연구)

  • Kim, Ye-Tae;Jin, Su-Eon;Kim, Hyun-Ki;Shin, Baek-Ki;Jeong, Ui-Hyeon;Kim, Chong-Kook;Park, Jeong-Sook
    • Journal of Pharmaceutical Investigation
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    • v.39 no.4
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    • pp.309-314
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    • 2009
  • An enzyme immunoassay (EIA) was validated for quantitation of cacitriol in human plasma. Calcitriol was immunoextracted with immunocapsules, which contain monoclonal antibodies to calcitriol linked to solid phase particles in suspension with a vitamin D binding protein inhibitor. Calcitrol was eluated and the eluates were evaporated under a gentle stream of nitrogen gas. The absorbance of analytes was determined using a microplate reader (reference wavelength 650 nm; measurement wavelength 450 nm). The method was specific and sensitive enough to detect as low as 6.5 pmol/L of calcitriol. Linear calibration range was 6.5-491 pmol/L with correlation coefficient greater than 0.99. The overall accuracy was in the range of 83.8 to 111.2% and precision C.V. (%) 0.99 to 8.47%. The recovery was approximately 100% and stability was confirmed during storage and sample preparation. The pharmacokinetic parameters were calculated by baseline subtraction because calcitriol is an endogenous material. Following oral dose of calcitriol, the mean AUC$_{24h}$ was 1038${\pm}$539 pmol/Lhr and C$_{max}$ of 128${\pm}$63.1 pmol/L was reached at 3.50${\pm}$1.07 hr. The mean t$_{1/2}$ of calcitriol was 5.13${\pm}$2.10 hr. The present EIA method was successfully applied to study bioavailability after oral administration of 2 ${\mu}$g of calcitriol in healthy Korean subjects.