Detection of messenger RNA in routinely processed tissue sections with biotinylated oligonucleotide probes.
Detection of messenger RNA in routinely processed tissue sections with biotinylated oligonucleotide probes.
复制标题
使用生物素化寡核苷酸探针检测常规处理的组织切片中的信使 RNA。
DOI：
10.1093/ajcp/92.2.166
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发表时间：
1989
影响因子：
3.5
通讯作者：
Lloyd,RV
中科院分区：
文献类型：
--
作者：
Hankin,RC;Lloyd,RV
使用放射性 35 S 标记探针和人降钙素生物素化寡核苷酸探针进行原位杂交 (ISH)，用于分析 8 例甲状腺髓样癌 (MTC) 的石蜡切片。其中三个病例还用冷冻组织切片进行了研究。生物素化探针可在 24 小时内轻松检测到常规处理的福尔马林固定石蜡包埋组织切片中的降钙素信使 RNA (mRNA)。 Northern 杂交和其他对照研究证明了降钙素探针的特异性。针对其他高丰度 mRNA（如促肾上腺皮质激素 (ACTH)、催乳素和生长激素）的生物素化寡核苷酸探针也检测到垂体组织中各自的 mRNA。这些结果表明，生物素化寡核苷酸cDNA探针可用于通过ISH检测某些内分泌细胞和肿瘤中大量存在的特定mRNA。这种方法提供了一种替代方案，不需要使用分子克隆或放射性探针来进行这种研究和诊断技术。
In situhybridization (ISH) with a radioactively35S-labeled probe and a biotinylated oligonucleotide probe for human calcitonin was used to analyze eight cases of medullary thyroid carcinoma (MTC) in paraffin sections. Three of these cases were also studied with frozen tissue sections. The biotinylated probe readily detected calcitonin messenger RNA (mRNA) in routinely processed formalin-fixed paraffin-embedded tissue section within 24 hours. Northern hybridization and other control studies demonstrated the specificity of the calcitonin probe. Biotinylated oligonucleotide probes for other mRNAs present in high abundance such as adrenocorticotroipic hormone (ACTH), prolactin, and growth hormone also detected the respective mRNAs in pituitary tissues. These result show that biotinylated oligonucleotide cDNA probes can be used to detect specific mRNAs present in large amounts in some endocrine cells and tumors by ISH. This approach offers an alternative that does not require the use of molecular cloning or radioactive probes for this investigative and diagnostic technique.