可溶性FL酶联检测试剂盒的研制及其运用  

Preparation and Application of ELISA Assay for Detecting Human Soluble Antigen Flt3-Ligand

作　　者：蒋靓[1] 蒋玉平[2] 张世杰[2] 王凤鸣[2] 孙静[2] 顾宗江[2] 

机构地区：[1]苏州市红十字中心血站,江苏苏州215006 [2]苏州大学医学生物技术研究所,江苏苏州215007

出　　处：《苏州大学学报（医学版）》2010年第4期767-769,共3页Suzhou University Journal of Medical Science

摘　　要：目的建立灵敏、特异和稳定的人可溶性FL(sFL)酶联检测试剂盒,探讨其临床检测意义。方法采用鼠抗人FL单克隆抗体2D2为包被抗体,识别人FL不同位点的商品化多克隆抗体为检测抗体,建立双抗体夹心的人sFL酶联免疫检测方法(sFL-ELISA)。在此基础上,分别对不同血清样本中sFL的含量进行定量分析。结果成功研制人sFL酶联检测试剂盒,灵敏度为7.8pg/ml。该试剂盒4℃放置3个月,离散度(CV%)<7.6%,加入rh-FL/Fc重组蛋白后的回收率在97%～110%。应用该试剂盒测得的健康供血员、再生障碍性贫血患者和白血病患者血清中sFL含量分别为(328.6±42.89)、(2607±286.8)、(1091±255.2)pg/ml。结论研制成灵敏、特异和稳定的人sFL酶联检测试剂盒,能对血清中sFL进行准确定量分析,具有良好的应用前景。Objective To establish a sensitive,stable ELISA assay for detecting human soluble antigen FL and exploring its clinical values.Methods The McAb 2D2 against human FL was used as coating antibody,the polyclonal antibody recognizing different epitope was used for testing antibody.Then a ELISA kit for detecting sFL was set up.The concentrations of soluble FL in serum samples were analyzed.Results ELISA method for measuring soluble human FL was successfully established.The detecting limit was 7.8 pg /ml.The CV of kit was less then ± 7.6% and the retrievable rate was 97% ～ 110% within three months as the kit was stored in 4℃.It was evidenced that the kit was of high sensitivity,stability and accutacy.The sFL concentrations in sera from healthy donors and patients with aplastic anemia and leukemia were（328.6 ± 42.89）,（2 607 ± 286.8） and（1 091 ± 255.2） pg /ml respectively.Conclusion Establishment of sFL detecting kit may be valuble in basic research,diagnosis,curative effect evaluation and prognosis estimation in sFL-related diseases.

关 键 词：可溶性FL 酶联免疫反应 白血病 再障 

分 类 号：R446.112[医药卫生—诊断学]