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目的发现和克服ＲＴ－ＰＣＲ中可能存在的假基因的干扰。方法采用ＲＴ－ＰＣＲ法扩增人肺鳞癌标本中角蛋白１９（ＣＫ１９ｃＤＮＡ或ＤＮＡ序列。结果通过设立分别以基因组ＤＮＡ和未经逆转录的ＲＮＡ为模板的对照可观察到ＣＫ１９假基因的干扰现象。用脱氧核糖核酸酶预处理ＲＮＡ标本或在设计错配引物的基础上选择合适的退火温度可消除干扰。结论在运用ＲＴ－ＰＣＲ法分析哺乳动物基因表达时常存在假基因的干扰，以上方法可有效地发现和克服这种干扰作用
Objective To discover and overcome the interference of possible pseudogenes in RT-PCR. Methods The cytokeratin 19 (CK19 cDNA or DNA sequence) was amplified by RT-PCR from human lung squamous cell carcinoma specimens.Results The interference of CK19 pseudogene was observed by setting up the control with genomic DNA and reverse transcriptase RNA as template RNA interference with deoxyribonuclease pretreatment of RNA samples or design of mismatch primers based on the selection of the appropriate annealing temperature can be eliminated.Conclusion The use of RT-PCR analysis of mammalian gene expression often exist pseudogenes interference, the above method can Effectively detect and overcome this interference