Detection Of Intron 22 Inversion And Intron 1 Inversion In Patients With Hemophilia A And Comparison Of Methods

Objective:Intron 22 inversion(Inv22)in patients with hemophilia A(HA)in Shanxi Province were detected by LD-PCR and RT-NPCR,and intron 1 inversion(Inv1)were detected by double-tube multiplex PCR.Understand the incidence of Inv22 and Inv1,and to compare the two methods in order to improve the detection methods.Methods:Selected patients with HA that registered in Shanxi Hemophilia Management Center,using long-distance polymerase chain reaction(LD-PCR)and reverse transcription nested polymerase chain reaction(RT-NPCR)to detect Inv22 respectively.Inv1 was detected by double-tube multiplex PCR method.Results:30 patients with HA(28.6%)were successfully amplified by LD-PCR in 105 patients,among which 16 cases(53.3%)were Inv22 positive.30 patients with HA patients(76.9%)were successfully amplified by RT-NPCR in 39 patients,and 9 cases of them(30.0%)were Inv22 positive.7 cases were detected by LD-PCR and RT-NPCR simultaneously.The results of two methods were consistent.Inv1 was detected by double-tube multiplex PCR,and all amplifications were successful.2(2.5%)were Inv1 positive in 77 patients.The cases diagnosed by our study group previously were chosen as positive control,and normal people were chosen as negative control.Conclusion:(1)The incidence of Inv22 and Inv1 with HA patients in Shanxi Province were basically consistent with domestic and foreign research reports.(2)At present,LD-PCR is one of the standard methods for detecting Inv22,but the amplification efficiency was low.The results of RT-NPCR and LD-PCR were in accordance with the same HA patients.But the amplification efficiency of RT-NPCR was higher,and it was time saving and more economical.It is likely to become a more efficient method in detecting Inv22 in future.