检测MBL EXON Ⅰ 52位密码子突变PCR-SSP方法的建立

Establishment of PCR-SSP for detecting point mutation at codon 52 of MBL Exon Ⅰ

目的 建立甘露糖结合凝集素 (MBL) 5 2位密码子基因突变的序列特异性引物聚合酶链反应 (PCR SSP)检测方法。方法 收集静脉血标本 ,提取染色体DNA ,根据MBL基因序列设计引物 ,PCR扩增。结果 扩增产物进行序列分析 ,与预期目的基因序列相同 ,检测灵敏度为 0 .8μg/ml。结论 该方法是一种特异性好、较为灵敏的检测方法。

Objective To establish the method for detecting the point mutation at codon 52 of MBL Exon Ⅰ,sequence specific primer polymerase chain reactions (PCR-SSP).Methods To collect venous blood specimen and extract DNA,PCR-SSP was established with self-designed primers according to MBL genomic sequence.Results PCR product was identical to the expected aim gene by sequencing.Detection sensitivity was 0.8μg/ml.Conclusion This method has high specificity,excellent reproducibility and more sensitivitity.