期刊
PRENATAL DIAGNOSIS
卷 21, 期 9, 页码 753-759出版社
WILEY-BLACKWELL
DOI: 10.1002/pd.170
关键词
thalassaemia; preimplantation genetic diagnosis (PGD); single cell fluorescent PCR; single gene disorder; single-stranded conformation polymorphism (SSCP)
Haemoglobinopathies including alpha- and beta -thalassaemia are the world's most common class of single gene disorder. Prenatal diagnosis (PND) for beta -thalassaemia has been proven to be an effective strategy for controlling the incidence of new cases and is widely used in several countries where the disease is common. Successful preimplantation genetic diagnosis (PGD) protocols for beta -thalassaemia have been introduced using restriction fragment length polymorphism (RFLP), single-stranded conformation polymorphism. (SSCP) and denaturing gradient gel electrophoresis (DGGE). However, contamination and allele dropout (ADO) remain an important concern for all of these strategies. In the present study two PGD protocols for detecting beta -thalassaemia mutations (codon 41-42 and IVSI-110) and one for alpha -thalassaemia (SEA mutation) have been designed and tested. These methods contain failsafe mechanisms to reduce the risk of misdiagnosis due to ADO or contamination and utilise multiplex fluorescent PCR (F-PCR). Interestingly, amplification efficiency and ADO were significantly affected by the choice of DNA polymerase and the freshness of the single cells used. The close similarity between the DNA sequences of beta -globin and delta -globin was also found to be an important issue that necessitated careful design of primers for the beta -globin gene. Copyright (C) 2001 John Wiley & Sons, Ltd.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据