4种罕见β-地中海贫血基因检测膜条的制备及其临床应用

Preparation and clinical application of membrane strips for four rare gene mutation loci of β-thalassemia

目的建立4个β-地中海贫血基因罕见突变位点反向杂交膜条的基因诊断方法,以用于临床该疾病罕见样本的检测。方法设计并合成8条探针用于检测在中国人中发现的4个β-地中海贫血基因罕见突变位点,将探针点至醋酸纤维膜上,制成反向杂交膜条;通过检测4种罕见临床样本和30份临床阴性标本,对其临床检测效果进行评价。结果共制备了含8条寡核苷酸探针的反向杂交膜条,临床验证结果表明,均能检测出这4种罕见的基因突变位点,而阴性标本未检测出。结论研制的针对4个β-地中海贫血基因罕见突变位点反向杂交膜条敏感度高,特异性好,能准确检测4个β-地中海贫血基因罕见突变的标本,可临床推广应用。

Objective To establish gene diagnostic method of reverse hybridization membrane strips for four rare gene mutation loci of β-thalassemia, apply the method in detection of rare cases of β-thalassemia in clinic. Methods Eight probes were designed and synthesized to detect four rare gene mutation loci of β-thalassemia found in Chinese, the probes were dotted onto cellulose acetate membrane to prepare reverse hybridization membrane strips; the clinical detection effect was evaluated by detecting four rare clinical samples and thirty clinical negative samples. Results Reverse hybridization membrane strips containing eight oligonucleotide probes were prepared successfully, clinical verification showed that all the four rare gene mutation loci were detected, while no locus was detected in negative samples. Conclusion These reverse hybridization membrane strips targeting to four rare gene mutation loci of β-thalassemia has high sensitivity and specificity, which is worthy of clinical promotion and application.