in the present study,9 exon-containing DNA segments of dystrophin gene with 9 sets of oligonucleotide primers by two-step multiplex polymerase chain reaction (mPCR) were amplified. Subsequently,gene analysis was perfo...in the present study,9 exon-containing DNA segments of dystrophin gene with 9 sets of oligonucleotide primers by two-step multiplex polymerase chain reaction (mPCR) were amplified. Subsequently,gene analysis was performed in 36 cases of Duchenne mascular dystroply (DMD) and 4 cases of Becker muscular dystrophy(BMD). The findings showed that 17 cases of deletion were detected by using the first 5 sets of primers with a relatively high incidence of deletion detection and 2 more cases of deletion were detected by using the remaining 4 sets of primers. The total deletion rate detected by mPCR with 9 cases of primers was 47. 5% of the patients examined,suggesting that about 79. 1% of the patients with gene deletion could be detected. Thus,as a preliminary screening, the two-step mPCR can be used in the gene diagnosis of DMD/BMD. The method is not only simple, convenient and rapid,but also free from radiosotope trouble.展开更多
Objective To type haplotypes among the patients, carriers and normal offspring in a family of males with Becker muscular dystrophy in one generation by allelic fragment length polymorphism analysis. Methods Deletio...Objective To type haplotypes among the patients, carriers and normal offspring in a family of males with Becker muscular dystrophy in one generation by allelic fragment length polymorphism analysis. Methods Deletion analysis of the patients were performed using multiplex polymerase chain reaction (PCR) of amplification with 9 dystrophin exon primers. Intragenic short tandem repeat (STR) sequence (STR44, STR45, STR49 and STR50) were amplified by PCR to analyse allelic fragment length polymorphisms in the members of the family. Results The deletions of exons 17, 19 and 45, as well as deletions of allelic fragments at the loci of STR44 and STR45 were determined in the patients. Hemizygosity at those two loci were detected and carrier status ascertained in the mother of the patients. The normal haplotypes were typed in the sister of the patients. Conclusion The method of STR sequence polymorphism analysis can determine haplotypes at normal status or at risk status. It would be used in prenatal diagnosis and carrier detection in the families of Duchenne and Becker muscular dystrophy.展开更多
文摘in the present study,9 exon-containing DNA segments of dystrophin gene with 9 sets of oligonucleotide primers by two-step multiplex polymerase chain reaction (mPCR) were amplified. Subsequently,gene analysis was performed in 36 cases of Duchenne mascular dystroply (DMD) and 4 cases of Becker muscular dystrophy(BMD). The findings showed that 17 cases of deletion were detected by using the first 5 sets of primers with a relatively high incidence of deletion detection and 2 more cases of deletion were detected by using the remaining 4 sets of primers. The total deletion rate detected by mPCR with 9 cases of primers was 47. 5% of the patients examined,suggesting that about 79. 1% of the patients with gene deletion could be detected. Thus,as a preliminary screening, the two-step mPCR can be used in the gene diagnosis of DMD/BMD. The method is not only simple, convenient and rapid,but also free from radiosotope trouble.
文摘Objective To type haplotypes among the patients, carriers and normal offspring in a family of males with Becker muscular dystrophy in one generation by allelic fragment length polymorphism analysis. Methods Deletion analysis of the patients were performed using multiplex polymerase chain reaction (PCR) of amplification with 9 dystrophin exon primers. Intragenic short tandem repeat (STR) sequence (STR44, STR45, STR49 and STR50) were amplified by PCR to analyse allelic fragment length polymorphisms in the members of the family. Results The deletions of exons 17, 19 and 45, as well as deletions of allelic fragments at the loci of STR44 and STR45 were determined in the patients. Hemizygosity at those two loci were detected and carrier status ascertained in the mother of the patients. The normal haplotypes were typed in the sister of the patients. Conclusion The method of STR sequence polymorphism analysis can determine haplotypes at normal status or at risk status. It would be used in prenatal diagnosis and carrier detection in the families of Duchenne and Becker muscular dystrophy.
