【摘要】 目的　为减少RNA提取过程中不同细胞和组织间的交叉污染,提高逆转录PCR(reversetranscriptionPCR,RTPCR)结果的特异性和可靠性。　方法　建立冰冻组织特定区域的RNA快速提取及RTPCR检测技术。首先,制备5μm和20μm的连续冰冻切片,前者用于常规组织学染色和针对靶基因产物的免疫组织化学染色。依观察结果,在20μm的切片上,分离某一特定区域做快速RNA提取和RTPCR分析。结果　该方法能在1小时内完成微量组织的RNA提取,并在6小时内获得RTPCR结果。与常规RNA提取法比较,可有效排除因细胞间污染所致的假阴性或假阳性结果。结论　该方法适用于在多种类型组织混杂的标本内检测某一特定细胞群体和各群体间的基因表达以及表达差异,是对现有RNA提取和RTPCR检测技术的重要补充。更多还原

【Abstract】 Objective To establish a method for quick RNA extraction from specified histological region in frozen sections so as to avoid cross RNA contamination among different types of cells/tissues and to achieve specific RT PCR amplification.Methods Prepare serial frozen sections 5μm in thickness for histological and immunocytochemical examination.Based on the results,make one or two pieces of 20μm section and select a specified region for quick RNA extraction and RT PCR analysis.Results\ RNA extraction can be finished within an hour and the data of RT PCR obtained within 6 hours.Besides the high performance,this method can efficiently exclude pseudo positive or pseudo negative results caused by inter cellular RNA contamination.Conclusion\ This method is an important complement to the current techniques for RNA extraction and RT PCR.更多还原