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目的探讨基于PCR结合反向斑点膜杂交技术(PCR-RDB)的地贫基因复合型检测方法的准确性、稳定性、临床应用优势及前景。方法收集165份已知不同地贫基因型和地贫阴性血液标本,采用基于PCR-RDB的地贫基因复合型检测方法进行检测;另外与现有临床地贫基因分开检测的方法进行对比实验,采用两种方法同时检测300例血液和干血斑标本。结果 165份血液标本的PCR-RDB复合型检测方法的结果和已知基因型完全一致。两种方法对比实验的检测结果也完全一致。300份临床标本中检测结果为:地贫阴性标本235例,占78.3%;地贫阳性标本65例,占21.7%,阳性标本中缺失型α地贫33例,非缺失型α地贫2例,β地贫25例,α地贫复合β地贫标本5例。结论基于PCR-RDB的地贫基因复合型检测方法可以准确、稳定性地进行地贫缺失型突变和点突变的检测,具有实验操作更加简捷和方便等优点,并提高了检测通量,在临床和地贫防控工作中将得到广泛的推广应用。
OBJECTIVE: To investigate the accuracy, stability, clinical advantages and prospect of thalassemia multiplex detection based on PCR combined with reverse dot blot hybridization (PCR-RDB). Methods Totally 165 blood samples of known thalassemia and thalassemia were collected and detected by multiplex PCR based on PCR-RDB. In addition, the method was compared with the existing methods for detecting gene of thalassemia. Two methods were used to simultaneously detect 300 blood and dried blood spots. Results The results of PCR-RDB multiplex assay in 165 blood samples were completely consistent with the known genotypes. The test results of the two methods are exactly the same. The test results in 300 clinical specimens were as follows: 235 cases of thalassemia negative samples (78.3%), 65 cases of thalassemia positive samples (21.7%), 33 cases of deletional α-thalassemia positive samples and 2 cases of non-deletional α-thalassemia , 25 cases of β thalassemia, and 5 cases of α thalassemia complicated with β thalassemia. Conclusion The detection of thalassemia gene complex based on PCR-RDB can accurately and stably detect the deletion mutation and point mutation of thalassemia. It has the advantages of simple and convenient experimental operation and higher detection flux. In the clinical And prevention and control of thalassemia will be widely promoted.