PCR-金磁微粒层析法在叶酸代谢相关基因检测中的应用研究-东南大学学报医学版

PCR-金磁微粒层析法在叶酸代谢相关基因检测中的应用研究

单位：连云港市妇幼保健院遗传实验室, 江苏连云港 222000

关键词：PCR-金磁微粒层析法 5 10-亚甲基四氢叶酸还原酶甲硫氨酸合成酶还原酶

分类号：R446.7

出版年·卷·期（页码）：2019·38·第五期（890-895）

摘要：

目的：对PCR-金磁微粒层析法检测人叶酸代谢相关基因进行方法学评价，并初步应用于临床检测。方法：同时应用PCR-金磁微粒层析法和Sanger测序法两种方法分别检测221例临床样本的5，10-亚甲基四氢叶酸还原酶（MTHFR）C677T、A1298C位点及甲硫氨酸合成酶还原酶（MTRR）A66G位点的基因型，并对两种方法检测的结果进行比较，以评估PCR-金磁微粒层析法的准确性、灵敏度及特异性。结果：两种方法检测同一批样本的结果完全一致，准确性达100%。PCR-金磁微粒层析法的灵敏度高，对MTHFR C677T、A1298C位点的最低检测限分别为2.5、1.25 ng·μl^-1，MTRR A66G位点的最低检测限为1.25 ng·μl^-1；特异性强，扩增试剂的扩增反应呈高度专一性。结论：PCR-金磁微粒层析法检测人叶酸代谢相关基因快速、简便，结果准确，适合应用于各级具有PCR实验室资质的医疗机构。

Objective: To evaluate the method of PCR-gold magnetic particle chromatography for detecting the folic acid metabolism-related genes, and initially apply it to clinical testing. Methods: The genotypes of 5,10-methylenetetrahydrofolate reductase (MTHFR C677T,A1298C) and methionine synthetase reductase (MTRR A66G) from 221clinical samples were detemined by the method of PCR-gold magnetic particle chromatography and Sanger sequencing, and then the results of two different detection methods were compared to evaluate the accuracy, sensitivity and specificity of PCR-gold magnetic particle chromatography. Results: The results of testing from the same batch detected by two ways were completely consistent with 100% accuracy. The sensitivity of PCR-gold magnetic particle chromatography was high, the minimum detection limits for MTHFR C677T and A1298C sites were 2.5 ng·μl^-1 and 1.25 ng·μl^-1, respectively, the minimum detection limit of MTRR A66G site was 1.25 ng·μl^-1; Besides, its specificity was strong and the amplification reaction was highly specific. Conclusion: PCR-gold magnetic particle chromatography for detecting the folic acid metabolism-related genes has many advantages, such as rapid analysis, easy operation, accurate and stable, suitable for large-scale clinical samples, which can be highly applicable to all levels of medical institutions with PCR lab.

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