中文摘要：

客观 OCT4A 被知道了在胚胎的干细胞的 pluripotency 的维护起一个关键作用。最近的研究证明了 OCT4A 也在部分肿瘤房间线和纸巾被表示。这研究被瞄准从 OCT4B， pseudogene，和 genomic 污染为 OCT4A mRNA 和辨别的相对量的察觉开发即时反向的 transcriptase 聚合酶链反应(RT-PCR ) 试金。锁的 nucleic 酸(LNA ) 修改了探查的方法被设计认出单个基础差别 352A/C 识别 OCT4A mRNA。一份 exon 连接教材被设计避免假积极由 genomic 污染引起了。另外，保留基因 glyceraldehyde-3-phosphate 脱氢酶(GAPDH ) 的一幢房子在平行被测量使样品和操作之间的差别正常化。结果实验证明最新确定的 RT-PCR 试金有选择地放大了 OCT4A mRNA；OCT4A 类似物给了否定信号。，房间线 nTERA-2 和 HepG2 为 HeLa 和 293 房间在 OCT4A 表示显示出积极结果线，以及主要的外部血 mononuclear 房间(PBMC ) ， OCT4A 表示是否定的。另外， OCT4A mRNA 的相对数量被周期阀值(Ct ) 计算方法和房子保留基因正规化。这种技术证明了为有高特性的 OCT4A mRNA 的相对 quantitation 有效的结论。

英文摘要：

Objective： OCT4A has been known to play a critical role in the maintenance of pluripotency of embryonic stem cells. Recent research has shown that OCT4A is also expressed in partial tumor cell lines and tissues. This study is aimed to develop a real-time reverse transcriptase polymerase chain reaction （RT-PCR） assay for relative quantitative detection of OCT4A mRNA and discrimination from OCT4B, pseudogene, and genomic contaminations. Methods： A locked nucleic acid （LNA）-modified probe was designed to discern the single base difference 352A/C to identify OCT4A mRNA. An exon-junction primer was designed to avoid false positive caused by genomic contaminations. In addition, a house keeping gene glyceraldehyde-3-phosphate dehydrogenase （GAPDH） was measured in parallel to normalize the differences between samples and operations. Results： Experiments showed that the newly established RT-PCR assay amplified the OCT4A mRNA selectively; OCT4A analogues gave negative signals. Cell lines nTERA-2 and HepG2 showed positive results in OCT4A expression, while for HeLa and 293 cell lines, as well as primary peripheral blood mononuclear cells （PBMCs）, OCT4A expression was negative. Additionally, the relative quantity of OCT4A mRNA was calculated by cycle threshold （Ct） method and house keeping gene normalization. Conclusions： This technique proved to be effective for relative quantitation of OCT4A mRNA with high specificity.