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改进的血浆中精氨酸加压素提取方法及放射免疫分析法:测定前样本处理的作用及儿童参考值

Improved extraction procedure and RIA for determination of arginine8-vasopressin in plasma: role of premeasurement sample treatment and reference values in children.

作者信息

Kluge M, Riedl S, Erhart-Hofmann B, Hartmann J, Waldhauser F

机构信息

Department of Pediatrics, University of Vienna, Währinger Gürtel 18-20, A-1090 Vienna, Austria.

出版信息

Clin Chem. 1999 Jan;45(1):98-103.

PMID:9895344
Abstract

We optimized an RIA for measurement of arginine8-vasopressin (AVP) in plasma by use of 100-mg Isolute C18 columns for extraction, addition of a preincubation step, and use of maximal dilution of a commercially available antiserum. The detection limit was 0.06 ng/L when 0.5 mL of acidified plasma was extracted. The within- and between-run CVs (n = 16) at physiological concentrations were 5.8-10. 2% and 6.5-11.7%, respectively. Prolonged storage of blood at 25 degreesC, but not at 4 degreesC, led to a significant increase in measured plasma AVP concentrations. When plasma samples were prepared at several centrifugation speeds, plasma AVP was significantly correlated with the platelet count (r = 0.899; P <0. 001). This emphasizes the need for careful sample preparation to avoid contamination of plasma with platelet-bound AVP. Basal plasma AVP in 203 children and adolescents (105 males and 98 females; ages, 1 day to 18 years) averaged 1.1 +/- 0.6 ng/L. There was no significant difference between males and females and no correlation with age. In 16 healthy adult controls, plasma AVP averaged 1.0 +/- 0.5 ng/L.

摘要

我们通过使用100毫克的Isolute C18柱进行萃取、增加预孵育步骤以及对市售抗血清进行最大程度稀释,优化了一种用于测量血浆中精氨酸8 - 血管加压素(AVP)的放射免疫分析方法。当萃取0.5毫升酸化血浆时,检测限为0.06纳克/升。在生理浓度下,批内和批间变异系数(n = 16)分别为5.8 - 10.2%和6.5 - 11.7%。在25℃而非4℃下长时间储存血液会导致测得的血浆AVP浓度显著升高。当以几种不同的离心速度制备血浆样本时,血浆AVP与血小板计数显著相关(r = 0.899;P < 0.001)。这强调了仔细制备样本以避免血浆被血小板结合的AVP污染的必要性。203名儿童和青少年(105名男性和98名女性;年龄从1天至18岁)的基础血浆AVP平均为1.1±0.6纳克/升。男性和女性之间无显著差异,且与年龄无关。在16名健康成人对照组中,血浆AVP平均为1.0±0.5纳克/升。

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