• 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.

• Korean Journal of Animal Reproduction
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• v.17 no.4
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• pp.347-356
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• 1994

A simpled and sensitive microplate enzyme immunoassay(EIA) was developed for the determination of progesterone concentration in serum, based on progesterone monoclonal antibody as anti-progesterone, horseradish peroxidase(HRP) as enzyme-label and tetramethylbenzidine(TMB) as substrate. The assay has a sensitivity of 5 pg-120pg/well and intra- and inter-assay coefficients of variation for progesterone standard curve (1.0ng~10.0ng/ml) were ranged 2.5~9.9% and 1.7.8.0%, respectively, determination coefficient of the regressio equation of our standard curve(R2=0.990$\pm$0.007) were high, and this is the same level as that of commercial kit(Hormonost Bio-Lab, Germany, R2=0.98~0.99). The progesterone concentration of serum determined by both kits (Work & Bio-Lab) were significantly correlated (r=0.95, P<0.01) although a little higher value were resulted in our kit than that of commercial kit. It generally is these results indicated that the microplate-EIA can be cused for the determination of progesterone in serum, as well as, for the determination of the early pregnancy diagnosis.

• Korean Journal of Veterinary Research
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• v.30 no.1
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• pp.35-40
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• 1990

This study was carried out to evaluate khe accuracy of the early diagnosis of pregnancy in dairy cows using EIK-kit of progesterone in milk and to compare the progesterone concentrations in milk with those assayed by radioimmunoassay. 1. The progesterone concentrations of the pregnant cows($2.07{\pm}0.54ng/ml$) were significantly higher than those of non-pregnant cows($1.04{\pm}0.19ng/ml$), and thereafter began to inerease and maintained high levels. 2. During 20 to 22 days after artificial insemination, the accuracy of pregnancy diagnosis from EIA-kit of progesterone were 93.7% for non-pregnant cows, and 91.6% for pregnant cows. 3. During 20 to 22 days after artificial insemination, the accuracy of pregnancy diagnosis from milk progesterone concentrations were 100% for non-pregnant cows (<2.7ng/ml), and 92.9% for pregnant cows($${\geq_-}3.2ng/ml$$). The average overall accuracy of prediction for pregnant and non-pregnant cows were 96.5%. 4. Accordingly, the pregnancy diagnosis from EIA-kit of progesterone is thought to be recommendable because this early diagnostic means are simple with accurate result.

• Korean Journal of Veterinary Research
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• v.31 no.2
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• pp.217-222
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• 1991

Most progesterone enzyme immunoassays(EIA) are used liquid phase double-antibody separation. These methods consume considerable time and reagents because of the requirements for several washing and centrifugation steps involving the reactants. Because of there several problems, we were prompted to develop an effective EIA system by the use of higher titer of progesterone antiserum free of anti-bovine serum albumin antibodies (anti-BSA). The results obtained were as follows. 1. The antibody of progesterone antiserum was high as $1.5{\times}10^5$. 2. Percent activity bound of progesterone antiserum was about 77 at a dilution to $5{\times}10^3$ times. 3. Progesterone antiserum was contained a large amount of anti-BSA antibodies. 4. The anti-BSA was completely absorbed by using of polymerised BSA. 5. The molecular weight of albumin polymer (polymerised BSA) obtained by using 2.5% glut. araldehyde was $5{\times}10^5$.

• Korean Journal of Animal Reproduction
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• v.13 no.3
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• pp.157-163
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• 1989

This stduy was carried out to evaluate the ability of clinical application of pregnancy diagnosis based upon the determination of progesterone in serum, utilizing EIA-kit of progesterone concentrations in the serum were assayed by radioimmunoassay. 1. The progesterone concentrations of the pregnant cows(2.40$\pm$0.34ng/ml) were significantly higher than those of non-pregnant cows(1.03$\pm$0.09ng/ml), and thereafter began to increase and maintained high levels. 2. During 20 to 22 days after artificial insemination, the accuracy of pregnancy diagnosis from EIA-kit of progesterone were 95.0% for non-pregnant cows, and 92.3% for pregnant cows. 3. During 20 to 22 days after artificial insemination, the accuracy of pregnancy diagnosis from serum progesterone concentrations were 100% for non-pregnant cows(<1.4ng/ml), and 96.2% for pregnant cows( 2.0ng/ml). The average overall accuracy of prediction for pregnant and non-pregnant cows were 98.1%. 4. Accordingly, the pregnancy diagnosis from EIA-kit of progesterone is thought to be recommendable because this early diagnostic means are simple with accurate results.

• 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.

• Korean Journal of Animal Reproduction
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• v.14 no.3
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• pp.165-173
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• 1990

A simple and sensitive microplate enzyme immunoassay(ELISA) was developed for progesterone, based on progesterone monoclonal antibody as anti-progesterone, horseradish peroxise(HRP) as enzyme-label and tetramethylbenzidine(TMB) as substrate. The assay has a sensitivity of 5pg-120pg/well and intra- and inter assay coefficients of variation for progesterone standard curve(0.1ng-3.2ng/ml) were ranged 4.4-10.6% and 5.-12.6%, respectively. They assay is performed in less than two hours and provide reliable values to differentiate among samples from day 0(A.I.), day 14 and day 19. The discriminatory levels for early pregnancy diagnosis are [>10ng(day 19) & decreasing rate <1.5 : pregnancy] and [ 7ng & decreasing rate 1.5 : non-pregnancy]. The accuracy of the pregnancy diagnosis for cows classified as positive(pregnancy) and negative(non-pregnancy) were 96% and 100%, respectively.

• Korean Journal of Animal Reproduction
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• v.13 no.3
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• pp.149-156
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• 1989

These experiment was carried out to improve the precision of early pregnancy diagnosis in dairy cattle. Changes in progesterone concentration of milk were measured by Enzyme Immunoassay(EIA) in 73 cows up to 21 days after insemination. The average concentraton of progesterone in milk was 1.9ng/ml at eatrus ; it increased to 17.8ng/ml on day 14, and thereafter it declined to 4.3ng/ml on day 21 in nonpregnant cows. Whereas in pregnant animals, it was maintained and elevated further to 22.2ng/ml on day 21. The accuracy of the pregnancy diagnosis for animals classified as positive (pregnant ; over 10ng/ml and decreasing rate<1.5) and negative (non-pregnant ; under 7ng/ml and decreasing rate>1.5) were 95% and 100% respectively. The samples ranging between 7ng/ml and 10ng/ml were classified as positive (decreasing rate<1.5) and negative(>1.5), which accuracy was 54.5% and 100% respectively. However this range appears to be of the most interest for the veterinary practitioner since cows in proestrus or early interestrus tend to have milk progesterone levels within this value. Causes for the insufficient precision of false pregnancy diagnosis are discussed.

• 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.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.169-177
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• 1995

This study was carried out to investigate the effect, range of practice, and propriety for diagnosis of early non-pregnancies and reproductive disorders by dairy cows' milk progesterone analysis used EIA-kit of home products. The results were summarized as follows : 1. During 2 to 6 months after artificial insemination, the results of milk progesterone measurement by Home-kit and Auto ELISA reader-kit with pregnant dairy cows(152 heads) certified by rectal palpation were revealed, in Home-kit, 145 heads(95.4%) of positive reaction, 7 heads(4.6%) of quasi-positive, and 0 heads(0%) of negative among 152 heads and, in Auto ELISA reader-kit, 152 heads(100%) of positive reaction among 152 heads. 2. During 19 to 22 days after artificial insemination, the results of milk progesterone measurement by Home-kit, and thereafrer during 50 to 90 days after that, the results of pregnant test by rectal palpation were summarized as follows : 147 heads(82.1%) among 179 heads of positive reaction by Home-kit and 5 heads(31.3%) among 16 heads of quasi-positive were revealed pregnant cows by rectal palpation, and 42 heads(100%) among 42 heads of negative were non-pregnant. 3. During 19 to 22 days after artificial insemination, the results of milk progesterone measurement by Auto ELISA reader-kit, and thereafrer during 50 to 90 days after that, the results of pregnant test by rectal palpation were summarized as follows : 146 heads(86.9%) among 168 heads of positive reaction by Auto ELISA reader-kit and 6 heads(28.6%) among 21 heads of quasi-positive were revealed pregnant cows by rectal palpation, and 48 heads (100%) among 48 heads of negative were non-pregnant. 4. For the accuracy of the rectal palpation, Home-kit and Auto ELISA reader-kit were used in the cows of ovarian diseases. The results were following : in the cows of reproductive disorders expected negative milk progesterone, the accuracies of rectal palpation were the same 75.5%(40 heads among 53 heads) by Home-kit and Auto ELISA reader-kit, and in the cows of reproductive disorders expected positive milk progesterone, the accuracies of rectal palpation were 82.6%(19 heads among 23 heads) and 91.3%(21 heads among 23 heads) by Home-kit and Auto ELISA reader-kit, respectively, and the general accuracies of rectal palpation were 77.6%(59 heads among 76 heads) and 80.3%(61 heads among 76 heads) by Home-kit and Auto ELISA reader-kit, respectively.