摘要
本文建立了高灵敏度C-反应蛋白的酶免疫分析(hs-CRP EIA或hs-CRP ELISA)方法。取抗CRP单克隆抗体C1在0.05mol/L pH9.0的碳酸缓冲液中将其包被在板条微孔内,成为固相抗体,以过碘酸钠方法将抗CRP单抗E11与辣根过氧化物酶相联结,成为标记抗体,与不同量的CRP标准品在磷酸缓冲液中保温反应,完成标准曲线。Bmax/B0为61.5,非特异结合小于1.6%,灵敏度小于0.1mg/L,批内CV为5.8%,批间CV为11.5%,回收率为98.8%。测定正常人450名:包括50岁以下270名,其中男160名、女110名,血清hs-CRP分别为1.34±0.80mg/L和1.25±0.92mg/L;50岁以上180名,其中男96名、女84名,hs-CRP分别为2.23±1.21mg/L和2.01±1.45mg/L。50岁以上组,血清hs-CRP值显著高于50岁以下组(P<0.01)。测定急性心肌梗死12例,血清hs-CRP为51.6±20.1mg/L,显著高于正常人组(P<0.0001),其他心血管疾病14例,hs-CRP也都显著高于正常人组(P<0.001),肺癌10例,hs-CRP为18.2±8.34mg/L,显著高于正常人组(P<0.001),其他肿瘤14例,hs-CRP也都显著高于正常人组(P<0.001)。新建hs-CRP ELISA与现行hs-CRP IRMA具有良好的相关性:Y=0.9198X,r2=0.9186。
The hs-CRP enzyme immunoassay was developed. Anti-CRP McAb Cl in 0.05 mol/L pH 9.0 borate buffer was coated on micro-wells of strips as solid phase antibody. Anti-CRP McAb Ell was labeled with horseradish peroxidase by using NaIO4 method. The standard curve was made by incubation of antibodies with different concentrations CRP standards in phosphate buffer. The Bmax/Bo ratio was 67.5, nonspecific binding 〈1.5 %, detection sensitivity 〈0. lmg/L. The intra-assay and inter-assay CV were 5.8% and 11.5% respectively. The recovery was 98.8%. The serum hs-CRP levels of 450 normal individuals were detected by enzyme immunoassay. The results showed that the serum hs-CRP levels in 270 males and females below the age of 50 were 1.34±0.89mg/L and 1. 25±0. 92 mg/L respectively; those in 180 males and females beyond the age of 50 were 2.23± 1.21mg/L and 2.01± 1.45mg/L respectively. The serum hs-CRP levels in the group of the age beyond 50 were significantly higher than those in group below 50 （P〈0.01）. The serum hs-CRP in 12 patients with acute myocardial infarction （51.6±20. 1mg/L） was significantly higher than that in normal individuals （P〈 0. 0001）. The other 14 patients with different cardiovascular diseases also showed significantly higher serum hs-CRP compared to normal individuals （P〈0. 001）. The serum hs-CRP in 10 patients with lung cancer （18. 2±8.34mg/L） was significantly higher than that in normal individuals （P〈0. 001）. The other 14 cases with different carcinomas also showed significantly higher serum hs-CRP compared to normal individuals （P〈0. 001）. The detected results with this new hs-CRP ELISA closely corresponded to those of detection with hs-CRP IRMA.
出处
《标记免疫分析与临床》
CAS
2008年第1期42-46,共5页
Labeled Immunoassays and Clinical Medicine
