摘要
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。
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 .
出处
《检验医学与临床》
CAS
2016年第18期2567-2568,2572,共3页
Laboratory Medicine and Clinic
基金
中南大学中央高校基本科研业务费专项资金资助项目(2015zzts304)
关键词
高分辨率熔解
ABO
血型
单核苷酸多态性
测序
high resolution melting
ABO blood type
single nucleotide polymorphism
sequencing
