Andresa Cardoso Grandini Almeida, Gabriela Pasqualim, Fabiana Q Mayer, Ida Vanessa Doderlein Schwartz, Carolina F Souza, Roberto Giugliani, Ursula Matte
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引用次数: 3
摘要
无义介导的衰变(NMD)是一种识别和降解含有过早停止密码子的信使RNA的机制。无义突变是导致i型粘多糖病的主要突变。为了确定NMD对基于cDNA测序的正确基因分型的影响,我们标准化了α - l -伊杜糖醛酸酶基因cDNA分子的测序,并在一组先前通过DNA分析确定突变的患者中验证了这一过程。虽然整个基因可以通过5次聚合酶链反应扩增出来,但cDNA不适合用于分子分析,因为携带剪接位点和无义突变的患者无法进行基因分型。
Analysis of cDNA molecules is not suitable for the molecular diagnosis of Mucopolysaccharidosis type I.
Nonsense-mediated decay (NMD) is a mechanism of the recognition and degradation of messenger RNA containing a premature stop codon. Nonsense mutations are the main mutations that lead to Mucopolysaccharidosis type I. To determine the effect of NMD on correct genotyping based on cDNA sequencing, we standardized the sequencing from alpha-L-iduronidase gene cDNA molecules and validated this process for a group of patients whose mutations had been previously identified by DNA analysis. Although the whole gene could be amplified in 5 polymerase chain reactions, cDNA proved unsuitable for molecular analysis as patients bearing splice site and nonsense mutations were not genotyped.
期刊介绍:
Diagnostic Molecular Pathology focuses on providing clinical and academic pathologists with coverage of the latest molecular technologies, timely reviews of established techniques, and papers on the applications of these methods to all aspects of surgical pathology and laboratory medicine. It publishes original, peer-reviewed contributions on molecular probes for diagnosis, such as tumor suppressor genes, oncogenes, the polymerase chain reaction (PCR), and in situ hybridization. Articles demonstrate how these highly sensitive techniques can be applied for more accurate diagnosis.