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目的竞争ＰＣＲ可用于ｍＲＮＡ的定量测定，为了得到白血病中ｂｃｒ－ａｂｌｍＲＮＡ定量ＰＣＲ的竞争内标物。方法利用重组ＰＣＲ技术，实施对ｂ３ａ２型ｂｃｒ－ａｂｌｃＤＮＡ中一２７６ｂｐ片段的缺失法定点突变。结果经ＤＮＡ序列分析证实，重组后的ｃＤＮＡ片段与重组前相比，５′和３′端大部分序列相同，仅中间５５ｂｐ的部分序列缺失，同时引入１９ｂｐ外源ＤＮＡ片段，即净缺失３６ｂｐ，得到２４０ｂｐ的重组ＰＣＲ产物。较ｂ２ａ２型ｂｃｒ－ａｂｌｃＤＮＡ相应的２０１ｂｐ扩增片段长３９ｂｐ，使之适于作为ｂ３ａ２和ｂ２ａ２型两类ｂｃｒ－ａｂｌｍＲＮＡ定量ＰＣＲ的通用内标物。结论表明重组ＰＣＲ是一种获得靶基因的竞争性内标物的简便而可靠的方法
The purpose of competitive PCR can be used for quantitative determination of mRNA, in order to obtain competitive leukemia bcr-ablmRNA quantitative PCR internal standard. Methods Recombinant PCR technique was used to perform site-directed mutagenesis on a 276 bp fragment in b3a2-type bcr-ablcDNA. Results The DNA sequence analysis showed that most of the 5 ’and 3’ ends of the recombined cDNA fragments had the same sequence except that the 55 bp partial sequence was deleted. At the same time, a 19 bp foreign DNA fragment was introduced, ie, the net deletion was 36 bp. A 240 bp recombinant PCR product was obtained. The 201bp amplified fragment corresponding to b2a2-type bcr-ablcDNA was 39bp in length, making it suitable as a universal internal standard for quantitative PCR of bcr-abl mRNAs of b3a2 and b2a2 types. The conclusion suggests that recombinant PCR is a convenient and reliable method for obtaining competitive internal standards for target genes