基于PCR-金磁纳米微粒层析技术检测乙醛脱氢酶2基因多态性方法的建立及应用

Establishment and Application of the Method of Polymorphism of Aldehyde Dehydrogenase 2 Gene Based on PCR-gold Magnetic Nanoparticle Chromatography

目的基于"PCR-金磁纳米微粒层析法"建立用于口腔拭子样本类型的乙醛脱氢酶2(ALDH2)Glu504Lys基因分型检测技术。方法以口腔拭子为样本类型,利用ARMS-PCR技术,针对ALDH2 Glu504Lys基因设计特异性引物,在实验室PCR常规反应条件上以金磁纳米层析试纸条为检测平台对各项条件进行优化,以确定最优PCR-金磁微粒ALDH2Glu504Lys口腔拭子反应体系和PCR反应程序。收集60例口腔拭子样本,利用金磁纳米层析检测技术进行检测,并用测序结果进行验证。结果检测的60例口腔拭子样本中杂合突变型16例,野生型42例,纯合突变型2例,金磁微粒层析法检测基因分型结果与测序结果符合率100%。结论建立了基于金磁纳米微粒ALDH2 Glu504Lys口腔拭子基因检测技术,该方法具有快速、简便和准确的特点,迎合了当下POCT的发展趋势,值得临床推广和应用。

Objective To estabolish the ALDH2 GluS04Lys genotyping technique for oral swab sample type based on ＂PCR- gold magnetic nanoparticle chromatographyl＇. Methods According to the ALDH2 GluS04Lys gene the specific primers were designed by ARMS-PCR technique. Optimized the optimal ALDH2 Glu504Lys oral swab reaction system and PCR reaction procedures based on the routine reaction conditions and reaction procedure of laboratory PCR. 60 samples of oral swabs were collected and tested by gold magnetic nano-chromatography. The results were verified by sequencing. Results The detected 60 cases of oral swab samples contain 16 cases heterozygous mutation, wild type 42 cases, homozygous muta- tion 2 cases and then the results by Goldmag nanoparticle detection was perfectly matched with sequencing results. Conclu- sion The Goldmag nanoparticles chromatographic detection technique of ALDH2 Glu504Lys buccal swabs system was es- tablished. With accuracy, fastness, reliability,it is worthy of clinical promotion.