血管紧张素转换酶基因插入/缺失多态性检测方法的研究

目的 采用三条引物法进行力紧张素转换酶（ACE）基因分型检测，并与Rigat法进行比较，以探讨Rigat造成DD型分型的错判率，并用此方法检测了中国汉族人群中ACE基因I／D基因型的分布频率以及插入（I）／缺失（D）多态性与高血压病（EH）间的相关性。方法 EH患（EH组）206例，用上述两种方法进行ACE基因分型；正常血压组（NT组）156例以及核心家系25个共118例，三条引物法进行ACE基因分型。结果 EH组Rigat法得出的DD型比例较三条引物法高。与三条引物法相比较，Rigat法对DD型的错判率为13．04％。用三条引物法进行基因分型的结果如下：（1）在NT组，ACE各基因型的分布频率为Ⅱ型0．51，ID型0．41，DD型0．08。等位基因频率为：I0．71，D0．29。（2）25个EH家系所有成员的基因分型结果，完全符合孟德尔遗传规律。（3）EH与NT两组间基因型和等位基因频率无显性差异。结论 三条引物法1次完成ACE基因分型，有良好的特异性、准确性和重复性。

Typing of human ACE gene I/D polymorphism: comparison of Rigat method to triple primer method

Objective There is insertion/deletion(I/D) polymorphism in human angiotensin I-converting enzyme (ACE) gene. The standard method developed by Rigat might sometimes mistype ID heterozygotes as DD homozygotes. The purpose of this study was to determine the mistyping rate of DD genotype by Rigat method compared with triple primer method, the distribution of ACE genotypes in Chinese, and the association of the ACE gene I/D polymorphism with essential hypertension. Methods 206 patients with essential hypertension were genotyped by both Rigat method and triple primer method, 156 normotensive subjects and 25 nuclear families including 118 individuals were genotyped by triple primer method only. Results 1. The frequencies of DD genotype and D allele detected by Rigat method in 206 essential hypertensives were higher than that detected by triple primer method. When 23 DD individuals genotyped by Rigat method were retyped using triple primer method, 3 of them (13.04%) were retyped as ID. 2. The frequencies of II, ID and DD genotypes in 156 normotensives were 51%, 41% and 8%, respectively. The frequencies of I and D alleles were 71% and 29%, respectively. 3. In 25 nuclear families, the ACE I/D polymorphism detected were well-accordant with Mendelian inheritance. 4. There was no significant difference in frequencies of both genotypes and alleles between hypertensive group and normotensive group. Conclusion Triple primer method could be specifically and accurately used to detect the ACE I/D polymorphism.