检测可溶性TREM-1的ELISA法的建立及初步应用

目的:建立定量检测可溶性TREM-1的抗体夹心ELISA法。方法:采用抗人TREM-1单克隆抗体(mAb)包被酶标板,以兔抗鼠TREM-1多克隆抗体为夹心抗体、HRP标记羊抗兔IgG为检测抗体、重组小鼠可溶性TREM-1为标准品,建立检测可溶性TREM-1的ELISA法,并对30例正常人和30例急性肺部感染患者血清样本进行了检测。结果:建立的夹心ELISA法检测TREM-1的线性范围为0.78～200μg/L,批内、批间变异系数分别为6.52%和9.46%。30例正常人和30例血清急性肺部感染患者TREM-1的含量分别为(0.69±0.18)μg/L和(1.16±0.42)μg/L,两者比较差异有统计学意义(P<0.001)。结论:成功建立了一种灵敏度高、稳定性好的检测可溶性TREM-1的抗体夹心ELISA法。

Establishment of ELISA for detection of soluble TREM-1 and preliminary application

Objective:To establish a ELISA sandwich method for quantitative detection of soluble TREM-1.Methods:Microtiter plates was coated with anti-human TREM-1 monoclonal antibody(mAb),rabbit anti-mouse TREM-1 polyclonal antibody was used for the sandwich antibody,HRP labeled goat anti-rabbit IgG was used for the detection of antibodies,recombinant mouse soluble TREM-1 was standard,and 30 normal subjects and 30 serum samples from patients with acute lung infection were tested.Results:By sandwich ELISA assay.TREM-1 in the linear range was 0.78~200 μg/L,intra and inter-coefficients of variation were 6.52% and 9.46%.the serum TREM-1 levels of 30 normal subjects and 30 patients with acute lung infection were(0.69±0.18) μg/L and(1.16±0.42) μg/L,the difference was statistically significant(P<0.001).Conclusion:A high sensitivity and good stability sandwich ELISA for detecting soluble TREM-1 is established.