摘要
目的采用一种新的铕螯合物BHHCT,制备高灵敏度荧光标记物,建立B型钠尿肽(BNP)固相时间分辨荧光免疫分析法(TrFIA).方法使用2株匹配的单克隆抗体,其中1株可以识别BNP分子C-端区,另1株生物素化单抗能识别分子内环结构,分别作为捕获抗体和检测抗体,并利用制备的SA-BHHCT-Eu3+作为示踪物,建立双抗体夹心法,检测正常对照组和心功能Ⅰ~Ⅳ级心力衰竭患者组血液BNP含量.结果本法测定BNP的灵敏度为12 pg/ml,批内和批间CV分别为3.9%和5.2%,平均回收率为97.3%,线性范围为20~4 000 pg/ml.对照组(n=110)、心功能Ⅰ级(n=31)、Ⅱ级(n=30)、Ⅲ~Ⅳ级(n=25)各组血浆BNP浓度分别为(69±37)、(84±46)、(346±134)和(932±326)pg/ml;心衰各组BNP水平显著高于对照组(P<0.05或P<0.001).以BNP 100pg/ml为cut off值,诊断心力衰竭(HF)的特异性为82.4%,敏感性为91.6%.结论应用铕螯合物BHHCT建立的固相TrFIA法测定BNP,是一个很好的诊断心衰和监测其严重程度的实验诊断指标.
Objeelive To develop a solid-phase time-resolved fluoroimmunoassay (TRFIA) with a new europium fluorescent chelate BHHCT for highly sensitive detection of B-type natriuretic peptide(BNP). Methods Two coupled monoclonal antibodies (MoAbs) to BNP32 were used,a two-side “sandwich” immunoassay which applied SA-BHHCT-Eu^3 ± as the marking substance to detect plasma BNP levels of healthy people( n = 110 ) and heart failure ( HF ) patients ( n = 86 ). Results The lowest detection level ( mean 3s ) was 12 pg/ml. The CVs of inter-and intra-assay were 3.9% and 5.2% , respectively. The mean recovery rate was 97.3%. The linear range of the method for BNP detection was 20 to 4000 pg/ml. The mean levels of plasma BNP concentration in healthy controls, HF patients ( with NYHA classification) class Ⅰ ( n = 31 ), class Ⅱ ( n = 30) and class Ⅲ - Ⅳ ( n = 25 ) were 69 ± 37 pg/ml, 84 ± 46 pg/ml, 336 -± 144 pg/m1,912 ± 379 pg/ml ,respectively ( P 〈0.05 or P 〈0. 001 ). BNP levels in the HF groups were significantly higher than that in healthy controls. Its sensitivity and specificity for the diagnosis of HF were 91% and 82.4% with a cut-off value of 100 pg/ml. Conclusions Our solid-phase TRIFA is a good method which can be used to detect the plasma BNP level of HF patients. Increased BNP indicates increment of the severity of HF according to the NYHA classification.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2005年第5期358-360,共3页
Chinese Journal of Clinical Laboratory Science
关键词
B型钠尿肽
时间分辨荧光免疫分析
铕螯合物
心力衰竭
B-type natriuretic peptide
time-resolved fluoroimmunoassay
europium-chelate
heart failure