新型亚甲基四氢叶酸还原酶单核苷酸多态性研究方法检测恶性血液病易感性

本研究探讨一种新型亚甲基四氢叶酸还原酶（MTHFR）单核苷酸多态性（SNP）检测方法，并用其检测恶性血液病遗传易感性。根据cDNA芯片原理制作一种目的基因芯片，利用双色荧光探针杂交进行SNP位点检测，测序法对该芯片检测结果的准确性进行验证，并以此对来自中国江苏地区的157例健康对照和127例恶性血液病患者（30例多发性骨髓瘤，28例非霍奇金淋巴瘤，22例急性淋巴细胞白血病，40例急性髓系白血病，7例慢性髓系白血病）的MTHFR基因C677T多态位点进行检测。结果表明，为野生型、杂合型和突变型的叠加荧光分别显示为绿色、黄色和红色。测序结果与芯片结果吻合。677C和677T在病例和对照组的基因频率分别为58．7％、66．9％、41、3％和33、1％，差异有显著性（χ^2=4．077，P=0．043）。677TT基因型发生MM相对风险明显增加（OR=4．21；95％CI=1．50-11．83；P=0．006）。结论：本芯片检测方法准确、高通量且价格低廉，适用于大规模样本SNP调查；C677T多态改变影响恶性血液病的发病风险。677TT基因型是MM的易感因素。

A New Method for 5,10-Methylenetetrahydrofolate Reductase Single Nucleotide Polymorphisms Genotyping Used to Study Susceptibility of Hematological Malignancy

The aim of this study was to set up a new method for 5, 10-Methylenetetrahydrofolate reductase (MTHFR) single nucleotide polymorphisms (SNP) genotyping, and to investigate the hereditary susceptibility of hematological malignancy. Prepared an aimed gene microarray based on cDNA microarray theory, dual-color fluorescence hybridization was used to detect SNP loci, and DNA sequencing was performed to confirm the results. The MTHFR C677T SNP loci of 157 controls and 127 patients with hematological malignancies (30 multiple myeloma, 28 non-Hodgkin's lymphoma, 22 acute lymphoblastic leukemia, 40 acute myeloid leukemia, 7 chronic myeloid leukemia) from Jiangsu province were detected. The results showed that after overlapping, homozygous wild type, heterozygote type and homozygous mutant type yielded green, yellow and red fluorescence, respectively. DNA sequencing validated these results. The allele frequency of 677C and 677T in patients and controls were 58.7% and 66.9%, 41.3% and 33.1% respectively, showing statistically significant difference (chi2 = 4.077, P = 0.043). 677TT genotype showed a significantly higher risk of MM (OR = 4.21; 95% CI = 1.50 - 11.83; P = 0.006). It is concluded that this microarray-based method is accurate, high-throughput and inexpensive, suitable for SNP genotyping in a large number of individuals. C677T polymorphisms influence the risk of hematological malignancies. 677TT genotype is susceptive to MM.