论文部分内容阅读
目的 建立一种检测先天性人巨细胞病毒感染胎鼠大脑皮质内皮素 1(ET 1)mRNA的方法。方法 采用逆转录 聚合酶链式反应 (RT PCR)方法并对实验条件进行优化。建立稳定的检测体系后 ,对胎鼠大脑皮质ET 1mRNA进行检测 ,并以RT PCR后产物的OD值来反映起初ET 1mR NA的模板相对含量。结果 在一定TaqDNA酶、镁离子浓度、引物浓度等条件下 ,ET 1mRNA经RT PCR扩增后产物的OD值与ET 1mRNA模板量成正比。结论 优化条件后的RT PCR是检测ET 1mRNA的一种敏感、简便、特异、可靠的方法。
Objective To establish a method for the detection of endothelin 1 (ET 1) mRNA in fetal rat cerebral cortex infected with congenital human cytomegalovirus. Methods Reverse transcription polymerase chain reaction (RT-PCR) was used to optimize the experimental conditions. After establishing a stable detection system, the ET 1mRNA of fetal rat cerebral cortex was detected, and the OD value of the product after RT PCR was used to reflect the initial relative content of ET 1mRNA. Results Under certain conditions of Taq DNA enzyme, Mg2 + concentration and primer concentration, the OD value of ET1mRNA amplified by RT PCR was proportional to the amount of ET1mRNA template. Conclusion RT PCR after optimized conditions is a sensitive, simple, specific and reliable method for detecting ET 1 mRNA.