预期应用

ELISA法定量测定人血清、血浆或其它相关液体中环胞霉素A(CsA)含量。

 

实验原理

环孢霉素A(cyclosporin A, CsA)系含11个氨基酸的环状多肽,是一种强效免疫抑制剂,对T淋巴细胞亚群具有选择性抑制作用,同时还抑制白细胞介素2、T细胞生长因子(TCGF)等细胞因子的释放。

本试剂盒应用双抗体夹心酶标免疫分析法测定标本中CSA水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入CSA抗原、生物素化的抗人CSA抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的CSA呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。

 

Intended use

This immunoassay kit allows for the specific measurement of Cyclosporin A（CsA） concentrations in cell culture supernates, serum and plasma.

 

Introduction

Cyclosporin A (CsA) has been reported to be effective in the treatment of nephrotic syndromecaused by minimal change disease and focal segmental glomerulosclerosis, and interest in small CSA for IMN has been aroused.Non-controlled small cohort studies have been published for CSA treatment of IMN, with a relatively high rate of remi ssion of nephrotic syndrome.Cyclosporine A (CsA) is an established drug for prevention of graft-versus-host-disease (GVHD) following hematopoietic stem cell transplantation and it can be administered intravenously or orally. It has been repeatedly shown that CsA binds to the inner surface of silastic, silicon or polyurethane central venous catheters.Blood collected from these catheters may result in falsely elevated drug levels. Therefore, where single-lumen catheters are present CsA drug levels should be taken from a peripheral vein, and in double-lumen catheters from the lumen not used for CsA infusion.

 

Test principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for CsA has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CsA present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for CsA is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of CsA bound in the initial step. The color development is stopped and the intensity of the color is measured.