应用PCR-RFLP技术进行ABO基因分型研究

Investigation into the genotyping of ABO blood group by PCR-RFLP

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。

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.