竞争性RT-PCR检测哮喘患者IL-8表达水平

Rapid Quantification of Interleukin-8 Gene Expression Level in Asthmatic Patients's Peripheral Blood by Competitive RT-PCR

利用反向ＰＣＲ法，缺失人ＩＬ－８ｃＤＮＡ内部ｂｐ序列，构建得到ＩＬ－８突变内标分子．用 已知量的该内标分子，通过竞争性ＲＴ－ＰＣＲ法、检测支气管哮喘患者外周血ＩＬ－８基因的表达情 况，发现患者ＩＬ－８ｍＲＮＡ的表达水平明显高于正常人对照组．这一结果初步表明，利用竞争 性ＲＴ－ＰＣＲ技术对支气管哮喘息者进行早期基因诊断，具有一定的可行性。

Competitive reverse tracscription polymerase chain reaction (competitive RT-PCR) is the most efficient method for accurately measuring mRNA level in samples of total RNA. We constructed mutant internal standards, which deleted 95 base pair nucleotides in internal coding sequence of interleukin-8 (IL-8) by inverse PCR technique. The expression level of IL-8 mRNA in asthmatic patients was determined by RT-PCR with this competitive internal standard. The experiment results showed that the mRNA level of IL-8 in asthmatic patients were markedly higher than that in normal people. This suggested it is possible for asthma early diagnoss using hte IL-8 internal standards.