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用聚合酶链反应（ＰＣＲ）方法制备地高辛精标记的单链探针，用以原位检测低丰度的β＿２肾上腺素能受体ｍＲＮＡ在大鼠肺组织中的分布。结果表明本方法制备的探针的检测敏感性显著高于用随机引物法标记的双链探针。已克隆于载体的ｃＤＮＡ或ＲＴ－ＰＣＲ产物均可作为模板合成单链探针，当ｄＡＴＰ、ｄＣＴＰ、ｄＧＴＰ浓度各为２００μｍｏｌ／Ｌ时，ｄＴＴＰ＋Ｄｉｇ－ｄＵＴＰ浓度降至５０～７５μｍｏｌ／Ｌ，扩增产量无显著变化，ｄＴＴＰ与Ｄｉｇ－ｄＵＴＰ浓度的适宜比例为３：１。本方法简便、稳定、敏感性高，可广泛用于原位检测低丰度ｍＲＮＡ。
Digoxigenin-labeled single-stranded probes were prepared by polymerase chain reaction (PCR) to detect the distribution of low abundance mRNA of β_2 adrenergic receptor in rat lung tissue. The results showed that the detection sensitivity of the probe prepared by the method was significantly higher than that of the double-stranded probe labeled with random primers. The cDNA or RT-PCR product cloned in the vector can be used as template to synthesize single-stranded probe. When dATP, dCTP and dGTP concentrations are 200μmol / L, the concentration of dTTP + Dig-dUTP is reduced to 50 ~ 75μmol / L, No significant change, dTTP and Dig-dUTP concentration of the appropriate ratio of 3: 1. The method is simple, stable and sensitive, and can be widely used for in situ detection of low abundance mRNAs.