科研之友
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摘要
The beta-thalassemias are genetic disorder caused by more than 200 mutations in the beta-globin gene, resulting in a total (beta(0)) or partial (beta(+)) deficit of the globin chain synthesis. The most frequent Mediterranean mutations for beta-thalassemia are: beta(0) 39, beta(+) IVSI-110, beta(+) IVSI-6 and beta(0) IVSI-1. Several molecular techniques for the detection of point mutations have been developed based on the amplification of the DNA target by polymerase chain reaction (PCR), but they could be labor-intensive and technically demanding. On the contrary, TaqMan (R) genotyping assays are a simple, sensitive and versatile method suitable for the single nucleotide polymorphism (SNP) genotyping affecting the human beta-globin gene. Four TaqMan r genotyping assays for the most common beta-thalassemia mutations present in the Mediterranean area were designed and validated for the genotype characterization of genomic DNA extracted from 94 subjects comprising 25 healthy donors, 33 healthy carriers and 36 beta-thalassemia patients. In addition, 15 specimens at late gestation (21-39 gestational weeks) and 11 at early gestation (5-18 gestational weeks) were collected from pregnant women, and circulating cell-free fetal DNAs were extracted and analyzed with these four genotyping assays. We developed four simple, inexpensive and versatile genotyping assays for the postnatal and prenatal identification of the thalassemia mutations beta(0) 39, beta(+) IVSI-110, beta(+) IVSI6, beta(0) IVSI-1. These genotyping assays are able to detect paternally inherited point mutations in the fetus and could be efficiently employed for non-invasive prenatal diagnosis of beta-globin gene mutations, starting from the 9 th gestational week.
- 出版日期2017-2-24
