标题：Establishment of a fluorescent PCR melting curve method for detecting asthma susceptibility using gene SNP typing
作者：Wang Na; Yang Rui-Rui; Chen Ming-Kai; Zhang Bin; Bie Qing-Li; Dong Feng-Yun; Liang Feng-Rong; Dong Hai-Xin; Wang Hai-Hua; Li Guang-T 更多 作者机构：[Wang Na; Chen Ming-Kai; Zhang Bin; Bie Qing-Li; Dong Feng-Yun; Dong Hai-Xin; Jin Cheng-Qiang] Jining Med Univ, Affiliated Hosp, Dept Clin Lab, Jining 更多
通讯作者：Jin, CQ;Jia, YX
通讯作者地址：[Jin, CQ]Jining Med Univ, Affiliated Hosp, Dept Clin Lab, Jining City 272029, Shandong, Peoples R China;[Jia, YX]Jining First Peoples Hosp, Radiol Dep 更多
来源：JOURNAL OF ASTHMA
关键词：Bronchial asthma; single nucleotide polymorphisms; genes; IL-13; IL-33;; GSDMA
摘要：Objective: To develop a detection method for single nucleotide polymorphisms (SNPs) of bronchial asthma (BA) susceptibility genes (IL-13, IL-33, and GSDMA) based on fluorescence PCR melting curves. Methods: Peripheral blood samples from 33 patients with BA were collected. DNA was extracted, and positive plasmids were constructed. Probes and primers for fluorescence polymerase chain reaction (PCR) were designed according to IL-13, IL-33, and GSDMA sequences, and the SNPs were separately detected by gene sequencing and fluorescence PCR melting curve. Results: The system was successfully divided into 3 SNPs, including IL-13, IL-33, and GSDMA, and a comparison of sequencing methods showed that the results were completely consistent. The lowest detection limit was 1 ng/reaction, the sensitivity and specificity were 100%, and this method had high repeatability (CV = 2.8%). Conclusion: The fluorescence PCR melting curve method is suitable for the rapid and accurate classification of SNPs. The method is economical, simple, and efficient, and is suitable for the screening of the susceptible gene SNPs in a large-scale population of patients with BA.