应用PCR快速制备乙型、丁型肝炎病毒诊断基因芯片探针

Rapid preparation of DNA microarray using PCR for hepatitis B and D virus detection

目的应用PCR技术扩增乙型、丁型肝炎病毒保守区域基因片段,并进行克隆、测序分析,制备联合诊断乙型、丁型肝炎病毒基因芯片探针。方法利用Oligo6.4软件分别针对乙型、丁型肝炎病毒基因保守区域设计PCR引物,纯化PCR扩增产物,扩增后的产物克隆至pMD18-T载体并进行快速鉴定,提取阳性克隆质粒进行测序分析及鉴定。结果获得多个乙型、丁型肝炎特异性基因片段。序列分析表明,所扩增的片段均属于乙型、丁型肝炎病毒特异基因。结论利用PCR扩增产物制备基因芯片探针是一种快速、简便的实用方法。

Objective To prepare DNA microarray for detecting both hepatitis B and D virus as (HBV and HDV). Methods With the assistance of Oligo6.4 software, specific PCR primers targeting the conserved region of HBV and HDV were de-signed. The PCR products were purified and cloned into the pMD18-T vectors, followed by rapid identification. The recombi-nant plasmids were then extracted from positive clones and the target gene fragments underwent sequence analysis. Results The gene fragments of HBV and HDV were obtained by PCR, which were confirmed by sequence analysis to be specific gene fragments of HBV and HDV. Conclusion Using PCR amplification products to prepare the DNA microarray is quick, simple and effective.