| 黄 嫄,兰雅娴,赵 雅,王 林,王晓春,马 虹.双向等位基因特异性PCR与限制性片段长度多态性基因分型方法比较[J].现代生物医学进展英文版,2017,17(28):5436-5440. |
| 双向等位基因特异性PCR与限制性片段长度多态性基因分型方法比较 |
| Comparison of Two Genotyping Methods: Bidirectinal PCR Amplification of Specific Alleles(Bi-PASA) and Restriction Fragment Length Polymorphism(RFLP) |
| Received:November 29, 2016 Revised:December 23, 2016 |
| DOI：10.13241/j.cnki.pmb.2017.28.008 |
| 中文关键词: 结直肠癌 EZH2 单核苷酸多态性 Bi-PASA PCR-RFLP |
| 英文关键词: Colorectal cancer EZH2 Single nucleotide polymorphism Bi-PASA PCR-RFLP |
| 基金项目:西安市科技计划项目（sF1516） |
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| 中文摘要: |
| 摘要 目的：比较双向等位基因特异性PCR（Bi-PASA）法与聚合酶链式反应-限制性片段长度多态性（RFLP）法对EZH2基因单核苷酸多态性（SNPs）位点rs887569基因分型结果有无差异，并用Bi-PASA法对EZH2基因rs17171119位点基因分型后分析与结直肠癌(CRC)易感性的相关性。方法：提取96名CRC患者与100名体检健康者的外周血DNA，分别用Bi-PASA法与聚合PCR-RFLP法检测EZH2基因单核苷酸多态性（SNPs）位点rs887569基因型，对两种分型结果进行比较；使用Bi-PASA法对EZH2基因rs17171119位点进行基因分型后用病例-对照方法分析该SNPs在中国人群中的分布。结果：Bi-PASA与PCR-RFLP对EZH2基因rs887569位点基因分型的准确率分别为99.5%和100%；EZH2基因的rs17171119 SNPs位点多态性与结直肠癌易感性无显著相关性（P=0.938,OR=0.846,95%CI:0.586-1.221）。结论：Bi-PASA是一种简单有效检测SNPs的方法，分型结果较为可靠；rs17171119 SNPs位点多态性与结直肠癌易感性无关，但本结论还有待更大样本量基因分型的验证。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate whether there are significant differences between genetyping results of EZH2 gene polymorphism rs887569 by bidirectinal PCR amplification of specific alleles (Bi-PASA) method and polymerase chain reaction- restriction fragment length polymorphism（PCR-RFLP） method respectively. And then we performed Bi-PASA on another SNPs (rs17171119) and investigate the association between SNPs(rs17171119) of EZH2 gene and colorectal cancer risk. Methods: We extracted the genomic DNA from the peripheral blood of 196 persons including 96 patients with CRC and 100 control participants. Then we performed EZH2 genotyping using Bi-PASA and PCR-RFLP respectively, compared the accuracy between the two kinds of genotyping methods. Then we performed Bi-PASA on another SNPs(rs17171119) of EZH2 gene and investigated the association between SNPs(rs17171119) of EZH2 gene and colorectal cancer risk. Results: The accuracy of Bi-PASA and PCR-RFLP are 99.5% and 100%, respectively. No statistically significant association was observed in rs17171119 and colorectal cancer risk（P=0.938, OR=0.846,95%CI:0.586-1.221）. Conclusion: Bi-PASA is a simple and effective method for detection of SNPs， its genotyping results are reliable. There was no evidence of any statistically significant correlation between rs17171119 of EZH2 gene polymorphisms and colorectal cancer risk. Nevertheless, further investigation with a larger sample size is needed to support our results. |
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