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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(χ2=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。
This study was to explore a novel method for the detection of MTHFR SNPs and to detect the genetic susceptibility to hematologic malignancies. According to the principle of cDNA microarray, a target gene chip was made, SNP locus was detected by double-color fluorescent probe hybridization, and the accuracy of the chip test results was verified by sequencing. Based on this, 157 healthy controls from Jiangsu Province, China MTHFR gene C677T polymorphism in 127 patients with hematologic malignancies (30 multiple myeloma, 28 non-Hodgkin’s lymphoma, 22 acute lymphoblastic leukemia, 40 acute myeloid leukemia and 7 chronic myeloid leukemia) Site for testing. The results showed that the superimposed fluorescence of the wild type, heterozygous type and mutant type showed green, yellow and red, respectively. Sequencing results coincide with the chip results. The frequencies of 677C and 677T in cases and controls were 58.7%, 66.9%, 41.3% and 33.1%, respectively. The difference was significant (χ2 = 4.077, P = 0.043). The relative risk of MM in the 677TT genotype was significantly increased (OR = 4.21; 95% CI = 1.50-11.83; P = 0.006). Conclusion The method is accurate, high-throughput and low-cost and suitable for large-sample SNP. The C677T polymorphism changes the risk of hematologic malignancies. The 677TT genotype is a predisposing factor for MM.