【摘要】 目的:探索一种优化的单细胞PCR佐剂,应用于分子病理学研究中单细胞基因的检测。方法:从淋巴结反应性增生冷冻组织切片中,应用激光捕获显微切割获取单个淋巴细胞,优化单细胞PCR反应体系进行actin基因检测。对照组不添加PCR佐剂,实验A、B、C组分别添加PCR佐剂二甲基亚砜、甘油、牛血清白蛋白。结果:对照组、实验A、B、C组的单细胞扩增阳性率分别为5%,7.5%,10%,45%,对照组和实验A、B组的单细胞扩增阳性率比较无显著性差异(P>0.05),而实验C组的单细胞扩增阳性率显著性高于对照组和实验A组(P<0.01)。结论:牛血清白蛋白是最佳的单细胞PCR佐剂,显著提高了单细胞PCR的敏感性、特异性和稳定性。更多还原

【Abstract】 Objective:To explore a kind of optimal PCR adjuvant for single cell gene detection in the study of molecular pathology.Methods:Single lymphocyte was microdissected from frozen tissue section of lymphnode with reactive hyperplasia,DNA from single cell was amplified with actin gene primers in optimized PCR system.Dimethyl sulfoxide,glycerine,bovine serum albumin were respectively added to the experimental A,B and C groups,while none of the PCR adjuvants were added to the control group.Results:The positive amplification rates were 5%,7.5%,10% and 45% in the control group,the experimental group A,B and C respectively,there were no significant differences between the control group and the experimental group A and B(P>0.05),while the positive amplification rate of the experimental group C was significantly higher than the other groups(P<0.01).Conclusion:Being optimal single cell PCR adjuvant,bovine serum albumin can effectively improve the sensitivity,specificity and consistency of single cell PCR.更多还原