摘要
目的:建立定量检测体液中微量水通道蛋白-2(AQP2)的酶联免疫吸附试验(ELISA),并用此方法检测羊水及新生儿尿液AQP2浓度。方法:人工合成AQP2蛋白特异性多肽片段,并与匙孔戚血蓝素联接制备兔抗AQP2多肽片段的多克隆抗体;建立检测AQP2的间接ELISA;并应用其对足月新生儿尿及孕足月羊水与成人尿样本进行AQP2的检测。结果:间接ELISA成功建立,灵敏度为2.5pmol/mL,批内及批间变异系数分别为4.15%和7.89%。用该法定量检测羊水与尿液中AQP2浓度,成人尿/新生儿3d尿中AQP2的浓度分别为(112.73±8.30)和(8.87±1.98)pmol/mL;孕足月羊水中未检测到AQP2的存在。结论:成功建立体液中微量AQP2间接ELISA,羊水中未检测到AQP2的表达。
Objective To develop a stable indirect enzyme-linked immunosorbent assay （ELISA） for detecting aquaporin-2 （AQP2） water channel protein in amniotic fluid and neonatal urine. Methods Human AQP2 specific peptides were synthesized and bound to KLH to prepare rabbit anti-AQP2 polyclonal antibodies. Amniotic fluid, neonatal and adult urine were collected to detect AQP2 protein by indirect ELISA. Results Indirect ELISA was successfully constructed to detect AQP2 as low as 2.5 pmol/mL with intra-and inter-assay coefficients of variance of 4.15% and 7.89%, respectively. The concentration of AQP2 protein was （112.73 ：t： 8.30）pmol/mL in adult urine and （8.87 ±1.98）pmol/mL in neonatal urine. However, AQP2 protein was undetectable in amniotic fluid. Conclusion Indirect ELISA has been successfully established to detect AQP2 in body fluid.
出处
《实用医学杂志》
CAS
2008年第8期1292-1294,共3页
The Journal of Practical Medicine
基金
广东省医学科学技术研究基金资助项目(编号:A2004542)
教育部留学回国人员科研启动基金资助项目(编号:教外司留[2005]383号)
关键词
水孔蛋白类
酶联免疫吸附测定
体液
Aquaporins
Enzyme-linked
immunosorbent assay
Body fluid
