一种简便的非放射性原位杂交法检测胰腺Ⅱ型弹性蛋白酶mRNA

A Convenient Non-radio ISH Method for Detection of Pancreatic Elastase Ⅱ mRNA

目的检测大鼠组织中胰腺Ⅱ型弹性蛋白酶mRNA的表达。方法根据大鼠胰腺Ⅱ型弹性蛋白酶基因序列合成两端含有相邻胸腺嘧啶结构的单链DNA探针,完成杂交后利用抗胸腺嘧啶二聚体抗体与抗原化的单链DNA探针进行反应,再运用免疫组织化学方法从而检出组织中目标mRNA的表达。结果大鼠胰腺组织的外分泌腺细胞的细胞质被特异性染色,其他的组织细胞均显示阴性。结论本试验中采用的原位杂交方法操作简便安全、无放射性,并且具有很高的灵敏度和特异性。实验结果提示大鼠胰腺外分泌腺细胞特异性表达胰腺Ⅱ型弹性蛋白酶mRNA,而其他组织细胞不表达。

Objective To detect the rat pancreatic elastase Ⅱ mRNA in rats.Methods ISH was preformed by using Oligo- DNA probe based on the sequence of rat pancreatic elastase Ⅱ mRNA, which 3＇ and 5＇ terminal conjugated with - A - T - T structure. The anti - T - T IgG antibody was used to react with haptenized Oligo - DNA in immunohistochemical process. Results Acinar cells in rat pancreas were highly labeled specifically by the probe. Conclusion It was a simple and safety ISH method with high sensitivity and specificity. The acinar cells in rat pancreas expressed pancreatic elastase Ⅱ mRNA specifically.