环孢素A放射受体测定方法的建立与考核

目的：建立用于临床的环孢素Ａ的放射受体测定法，并与荧光偏振免疫测定法进行比较。方法：利用亲环素Ａ（ＣｙＰＡ）能在体外与环孢素Ａ（ＣｓＡ）高效结合的特性，以重组人亲环素Ａ（ｒｈＣｙＰＡ）作为特异性的结合蛋白，３Ｈ－ＣｓＡ作为示踪剂，建立ＣｓＡ的放射受体测定法。结果：方法的有效测定范围为４３．８～２８００μｇ／Ｌ，灵敏度为４３．８μｇ／Ｌ，批内变异系数为８．２％（ｎ＝８），批间变异系数为１０．６％（ｎ＝８），标本提取与测定方法的全程回收率分别为１０４．２％和１０６．３％；对ＣｓＡ及其几种类似物与ｒｈＣｙＰＡ的亲和力进行比较，以ＣｓＡ与ｒｈＣｙＰＡ的亲和力为最佳；在实际应用中与荧光偏振免疫测定法进行比较（ｎ＝３０），二者测值高度相关，ｒ为０．９６４２，回归方程为ｙ＝３９．１１２＋０．９９ｘ。结论：该方法具有灵敏可靠，特异性、重复性、稳定性和经济性好等优点，适用于ＣｓＡ全血浓度的临床测定。

The radioreceptor assay of cyclosporine A and its identification

Objectives: To develop a radioreceptor assay of cyclosporine A( CsA ) in clinic, its results were compared with those from FPIA. Methods: Recombinant human cyclophilin A (rhCyPA) was applied as the binding protein with the use of 3 HCsA as tracer in the measuring procedure of CsA. Results: The assay could detect CsA at a range of 43.82 800 g/L.The sensitivity was 43.8 g/L. CVs were 8.2% for exterassay (n=8) and 10.6% for interassay ( n = 8) . The total recoveries of extraction procedure and analytical method were 104. 2% for 240 g/L and 106.3% for 764 g/L respectively, and the affinity of rhCyPA to CsA was better than those of CsA analogs( CsB,CsC, CsD ).The results obtained from the method were closely related with those from FPIA (n=30, r=0.9642, y=39.112+0.99x). Conclusions:] The radioreceptor assay of CsA is a sensitive, reproducible and stable method, it is useful for measuring CsA in whole blood.