Liang M H, Wong L J
Molecular Diagnostic Laboratory, Institute for Molecular and Human Genetics, Georgetown University, Washington, DC 20007, USA.
Am J Med Genet. 1998 Jun 5;77(5):395-400.
The multiplex polymerase chain reaction-allele specific oligonucleotides (PCR/ASO) dot blot hybridization method was used to detect 44 mitochondrial DNA point mutations in 2,000 patients suspected as having mitochondrial DNA disorders. These point mutations are classified into four categories. Category I consists of primary disease-causing, heteroplasmic point mutations. Homoplasmic nucleotide substitutions that have been reported to be possibly disease associated are in Category II. Homoplasmic nucleotide substitutions that are thought to be benign polymorphism are included in category III. The novel nucleotide substitutions recently discovered in our laboratory by single strand conformation polymorphism analysis are in category IV. Frequencies of these 44 nucleotide substitutions in 2,000 patients and 262 control individuals were studied. The results indicated that analysis of 12 recurrent disease-causing point mutations in category I identified 5.4% of the patients suspected as having mitochondrial DNA disorders. Since the mitochondrial disorders are a group of complex, heterogeneous, and multisystemic diseases, it is often difficult to confirm clinical diagnosis without molecular studies. Thus, the multiplex PCR/ASO method is an effective approach for initial screening of mtDNA mutations in patients suspected as having mitochondrial DNA disorders.
采用多重聚合酶链反应-等位基因特异性寡核苷酸（PCR/ASO）斑点杂交法，对2000例疑似线粒体DNA疾病的患者进行44种线粒体DNA点突变检测。这些点突变分为四类。第一类包括原发性致病异质性点突变。已报道的可能与疾病相关的同质性核苷酸替换属于第二类。被认为是良性多态性的同质性核苷酸替换包含在第三类。最近在我们实验室通过单链构象多态性分析发现的新型核苷酸替换属于第四类。研究了这44种核苷酸替换在2000例患者和262例对照个体中的频率。结果表明，对第一类中12种复发性致病点突变的分析，在疑似线粒体DNA疾病的患者中鉴定出5.4%。由于线粒体疾病是一组复杂、异质性和多系统性疾病，若无分子研究，往往难以确诊临床诊断。因此，多重PCR/ASO方法是对疑似线粒体DNA疾病患者进行mtDNA突变初步筛查的有效方法。
