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使用具有生物活性的重组火鸡催乳素开发和验证同源放射免疫测定法。

Development and validation of a homologous radioimmunoassay using a biologically active recombinant turkey prolactin.

作者信息

Guémené D, Bédécarrats G, Karatzas C N, Garreau-Mills M, Kuhnlein U, Crisóstomo-Pinto S, Zadworny D

机构信息

Station de Recherches Avicoles, INRA Centre de Tours, Nouzilly, France.

出版信息

Br Poult Sci. 1994 Dec;35(5):775-87. doi: 10.1080/00071669408417742.

DOI:10.1080/00071669408417742
PMID:7719741
Abstract
  1. A new homologous radioimmunoassay has been developed for the measurement of turkey prolactin. 2. A 25000 kDa purified recombinant derived turkey prolactin (rtPRL), the biological activity of which was tested using a crop sac assay, was used as immunogen for the production of rabbit antiserum. In this biological test, the rtPRL was as active as the ovinePRL. 3. The radioligand (rtPRL) was labelled with 125I and the assay allowed the detection of standard doses of rtPRL ranging from 400 pg/tube to 50 ng/tube. 4. No cross reaction with chicken luteinising hormone and recombinant chicken growth hormone was detected. 5. The within and between assay coefficients of variability were 5.0 +/- 2.7% and 16.3%, respectively. The overall mean recovery ratio was 1.01. 6. The dose-response curves obtained with serial dilution of plasma and pituitary from turkey hens at different physiological stages and from male turkeys were parallel to those obtained with standard rtPRL. 7. The measured concentration of prolactin was 5 times higher in plasma from incubating than laying turkey hens, and the pituitaries from incubating hens contained 2 and 4 times more prolactin than those of laying and out of lay hens or males, respectively. 8. To further assess the validity of the assay, we measured changes in plasma concentration of prolactin in turkeys following stimulation with chicken vasointestinal peptide (cVIP). A single injection of 1 or 10 micrograms/kg body weight of cVIP to laying hens produced a large and rapid increase in plasma prolactin. 9. This new radioimmunoassay appears to be high for the measurement of turkey prolactin.
摘要
  1. 已开发出一种用于测量火鸡催乳素的新型同源放射免疫分析方法。2. 一种25000 kDa的纯化重组火鸡催乳素(rtPRL),其生物活性通过嗉囊试验进行了测试,被用作免疫原以产生兔抗血清。在该生物试验中,rtPRL与绵羊催乳素活性相当。3. 放射性配体(rtPRL)用125I标记,该分析方法能够检测出每管400 pg至50 ng范围内的标准剂量rtPRL。4. 未检测到与鸡促黄体生成素和重组鸡生长激素的交叉反应。5. 分析方法内部和之间的变异系数分别为5.0 +/- 2.7%和16.3%。总体平均回收率为1.01。6. 用来自不同生理阶段的火鸡母鸡和雄火鸡的血浆及垂体进行系列稀释所获得的剂量 - 反应曲线与用标准rtPRL获得的曲线平行。7. 孵化期火鸡母鸡血浆中测得的催乳素浓度比产蛋期母鸡高5倍,孵化期母鸡垂体中含有的催乳素分别比产蛋期和休产期母鸡或雄火鸡的垂体多2倍和4倍。8. 为进一步评估该分析方法的有效性,我们测量了用火鸡血管活性肠肽(cVIP)刺激后火鸡血浆中催乳素浓度的变化。向产蛋母鸡单次注射1或10微克/千克体重的cVIP会使血浆催乳素大幅快速增加。9. 这种新的放射免疫分析方法对于测量火鸡催乳素似乎具有较高的准确性。

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Br Poult Sci. 1994 Dec;35(5):775-87. doi: 10.1080/00071669408417742.
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