高分辨率熔解曲线在ABO血型基因526位点单核苷酸多态性检测中的应用

摘要

Objective To establish a method for detecting single nucleotide polymorphism(SNP) of 526 locus in ABO gene by u‐sing high resolution melting(HRM ) curve .Methods Genomic DNA was extracted from blood samples of 35 individuals randomly , whose ABO blood types were going to be detected .The HRM method was then used to test the polymorphism at 526 gene locus of genomic DNA in 35 individuals .The accuracy of results was further verified by PCR‐DNA sequencing and ABO blood type serologi‐cal test .Results Among 35 samples ,13 cases were heterozygote and 22 cases were homozygote .Verified by sequencing and sero‐logical tests ,13 cases of G/C heterozygote included 8 cases of B blood type and 5 cases of AB blood type respectively ,while 22 cases of CC homozygote included 10 cases of A blood type and 12 cases of O blood type .The results of HRM analysis were consistent with the results of DNA sequencing and serological tests .Conclusion The HRM method is accurate ,convenient ,efficient and eco‐nomical ,which is expected to be a new method for genotyping ABO blood group .%目的：建立高分辨率熔解（HRM ）曲线检测 ABO 血型基因526位点单核苷酸多态性（SNP）的方法。方法随机提取35例待检血型标本的 DNA ，运用 HRM 曲线检测 ABO 血型基因526位点的 SNP ，通过 PCR‐DNA 测序以及血型血清学试验验证 HRM 检测结果的准确性。结果35例血型标本中，共检出526位点杂合子13例，纯合子22例。经测序分析和血清学试验验证13例 G／C 杂合中 B 型8例、AB 型5例，22例 CC 纯合子中 A 型10例，O 型12例。 HRM 法检测结果与测序结果及血清学结果相符。结论 HRM 法是一种低成本、简便、准确的 SNP 检测技术，有望成为临床 ABO 血型基因分型的新方法。