双抗体夹心免疫PCR－检测神经肽Y

DOUBLE ANTIBODY SANDWICH IM-PCR-DETECTING NPY

建立了一种双抗体夹心免疫ＰＣＲ方法。用神经肽Ｙ单克隆抗体包被；多克隆抗体作为夹心抗体；生物素标记的羊抗兔ＩｇＧ和游离的亲合素作为连接分子；生物素化的腺病毒六邻体基因重组质粒ＤＮＡ作为指示分子；用腺病毒六邻体基因的特异引物扩增指标分子。结果表明本实验方法检测神经肽Ｙ的敏感性可达５．０×１０^（－９）μｇ／ｍｌ，或５．０×１０^（－１０）ｇ／ｍｌ，比ＥＬＩＳＡ（０．７８μｇ／ｍｌ）敏感１．５６×１０~８或１．５６×１０~９倍。

In this paper,the double antibody sandwich IM-PCR was constructed for detecting NPY with monoclone and polyclone antibodies to NPY;Bio-sheep-anti-mouse IgG and free avidin as linking molecules,and Bio-DNA as mark molecule. The template was amplified by the primers of adenovirus hexon gene. The results showed that the sensitivity of IM-PCR for NPY was 5.0×10￣(-9)μg/ml or 5. 0×10￣(-10)μg/ml,and more sensitive 1.56×10￣8 or 1.56×10￣9 times than ELISA(0.78 μg/ml).