Broyles D L, Nielsen R G, Bussett E M, Lu W D, Mizrahi I A, Nunnelly P A, Ngo T A, Noell J, Christenson R H, Kress B C
Beckman Coulter, Inc., San Diego, CA 92196, USA.
Clin Chem. 1998 Oct;44(10):2139-47.
The performance characteristics of the Tandem-MP Ostase assay, a new microplate immunoassay for bone-specific alkaline phosphatase (bone ALP; EC 3.1.3.1) in human sera, are described. Bone ALP is bound to streptavidin-coated microwells by a single biotinylated anti-bone ALP monoclonal antibody. Antigen is detected by the addition of p-nitrophenyl phosphate. The assay is performed at room temperature in <90 min. Imprecision was 2.3-6.1% with a detection limit of 0.6 microg/L. Method comparison of bone ALP measurements with the Tandem-MP Ostase assay and the mass-based Tandem-R Ostase assay (n = 285) indicated regression statistics of Tandem-MP Ostase = 1.03 Tandem-R Ostase + 0.22 microg/L, S(y/x) = 4.0 microg/L, r = 0.97. Serum bone ALP values in apparently healthy men and in pre- and postmenopausal women were also similar between the two Ostase assay formats. Liver ALP reactivity determined using the slope and heat inactivation methods was similar in both Ostase assays. Liver ALP reactivity ranged from 3 microg/L (heat inactivation) to 6 microg/L (slope method) per 100 U/L of liver ALP activity, whereas bone ALP reactivity was 37 microg/L per 100 U/L of bone ALP activity, indicating a liver ALP relative reactivity of 8.1-16.2%. Similar results were obtained with the Alkphase-B bone ALP immunoassay. The Tandem-MP Ostase bone ALP assay demonstrated increased concentrations of serum bone ALP in conditions where bone metabolism is increased and showed a rapid, temporal decrease in serum bone ALP in Paget disease patients on bisphosphonate therapy. In conclusion, the Tandem-MP Ostase assay for serum bone ALP is a rapid, simple, robust nonisotopic alternative to the Tandem-R Ostase immunoradiometric assay that provides an accurate and sensitive assessment of bone turnover.
本文描述了一种用于检测人血清中骨特异性碱性磷酸酶（骨ALP；EC 3.1.3.1）的新型微孔板免疫分析方法——串联MP骨碱性磷酸酶分析的性能特征。骨ALP通过单一生物素化抗骨ALP单克隆抗体与链霉亲和素包被的微孔结合。通过添加对硝基苯磷酸来检测抗原。该分析在室温下进行，耗时不到90分钟。不精密度为2.3 - 6.1%，检测限为0.6μg/L。对串联MP骨碱性磷酸酶分析和基于质量的串联R骨碱性磷酸酶分析（n = 285）进行的骨ALP测量方法比较表明，串联MP骨碱性磷酸酶分析的回归统计量为：串联MP骨碱性磷酸酶分析 = 1.03×串联R骨碱性磷酸酶分析 + 0.22μg/L，S(y/x) = 4.0μg/L，r = 0.97。在两种骨碱性磷酸酶分析形式中，明显健康男性以及绝经前和绝经后女性的血清骨ALP值也相似。使用斜率法和热灭活法测定的肝脏ALP反应性在两种骨碱性磷酸酶分析中相似。每100 U/L肝脏ALP活性的肝脏ALP反应性范围为3μg/L（热灭活法）至6μg/L（斜率法），而每100 U/L骨ALP活性的骨ALP反应性为37μg/L，表明肝脏ALP相对反应性为8.1 - 16.2%。使用Alkphase - B骨ALP免疫分析也获得了类似结果。串联MP骨碱性磷酸酶分析显示，在骨代谢增加的情况下血清骨ALP浓度升高，并且在接受双膦酸盐治疗的佩吉特病患者中血清骨ALP迅速出现暂时性下降。总之，用于血清骨ALP的串联MP骨碱性磷酸酶分析是一种快速、简单、可靠的非同位素方法，可替代串联R骨碱性磷酸酶免疫放射分析，能对骨转换进行准确且灵敏的评估。
