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应用PCR-RFLP技术进行ABO基因分型研究
引用本文:周代锋,蔡望伟,王政,蒙晶,王小英. 应用PCR-RFLP技术进行ABO基因分型研究[J]. 中国热带医学, 2008, 8(8): 1280-1284
作者姓名:周代锋  蔡望伟  王政  蒙晶  王小英
作者单位:1. 海南医学院,海南,海口,571101
2. 海南医学院附属医院儿科,海南,海口,571101
基金项目:1、海南省教育厅资助课题 , 2、海南医学院苗國基金资助课题 , 3、海南省自煞科学基金立项课题
摘    要:目的建立PCR-RFLP技术鉴定ABO血型基因型的方法。方法应用PCR技术扩增ABO基因第六外显子上分别包含258位核苷酸和700位核苷酸的两个DNA片段,分别用KpnI和AluI酶解PCR扩增产物,电泳分离鉴定ABO血型基因型,同时对上述两种PCR扩增的DNA片段进行序列测定。结果用PCR-RFLP技术鉴定的ABO基因分型结果与PCR扩增产物的序列测定结果完全符合,可鉴定出AA、AB、AO、BB、BO、OO等6种ABO血型基因型。结论PCR-RFLP技术操作简便,重复性和稳定性好,可作为鉴定ABO血型基因型的可行方法。

关 键 词:聚合酶链反应-限制性片段长度多态性(PCR-RFLP)  ABO血型  ABO基因  基因分型

Investigation into the genotyping of ABO blood group by PCR-RFLP
Affiliation:ZHOU Dai - feng, CAI Wang - wei, WANG Zheng, et al. (1. Department of Biochemistry, Hainan Medical College, Haikou 571101, Hainan, P. R. China)
Abstract:Objective To establish a PCR-RFLP technique for investigation of the genotyping of ABO blood group. Methods Two fragments of the sixth exon which include the 258 locus and 700 locus were amplified separately by PCR method, then the PCR product was digested by KpnI and AluI and ABO blood type was anlayzed based on the results in the agarose gel. On the other hand, the ABO blood type was analyzed according to the sequence of the PCR product. Results The ABO blood type identified by PCR-RFLP methods is exactly the same to that identified by sequencing, both of which could i-dentify six ABO blood types (AA, AB, AO, BB, BO, OO). Conclusion PCR-RFLP with advantages of stability and simplicity is an effective method for investigation of ABO blood type.
Keywords:Polymerase chain reaction - restriction fragment length polymorphism ( PCR - RFLP)  ABO blood group  ABO gene  Genotyping
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