从OCT包埋冰冻组织提取组织RNA和检测病毒RNA

RNA extraction and virus RNA detection from frozen OCT- embedded tissue

目的探讨如何从OCT包埋冰冻组织提取组织RNA和检测病毒RNA。为临床有限标本的保存和利用提供一简便的方法。方法用汉坦病毒感染的小鼠组织标本,经质量浓度25%蔗糖磷酸缓冲液固定OCT包埋,在-20℃冰箱中保存2年后,对冰冻的OCT包埋的感染组织标本进行RNA提取,用凝胶电泳、紫外分光光度计及RT-PCR等方法检测组织总RNA和病毒RNA。结果组织经质量浓度25%蔗糖磷酸缓冲液固定OCT包埋冰冻保存后,从中提取的RNA仍保持较好的完整性,可扩增出较长片段的病毒RNA。结论OCT包埋的冰冻组织标本不影响提取组织RNA并检测病毒RNA。

Objective To study the approach of extracting RNA and detecting virus RNA from frozen OCT-embedded tissue. Methods The tissues sample of Hantaan virus infected mouse fixed with 25 % sucrose buffer solution and embedded with OCT,cryopreserved at -20 ℃ for 2 years. Then RNA was extracted from the embedded tissue and the total tissue RNA and viral RNA were detected by gel electrophoresis,ultraviolet spectrophotometry and RT-PCR. Results The RNA extracted from the frozen OCT-embeded tissue still remained better integrity,it could be amplified to longer segment of viral RNA. Conclusion It demonstrates that there is no influence on extracting tissue RNA and detecting viral RNA from frozen OCT-embedded tissue sample.