Abstract
目的 建立串联重复序列信号放大（TRSE）液相杂交系统检测HBV-DNA技术,并初步评价其效果.方法 在前期研究构建的含24个60 bp串联重复核酸片段的重组噬菌粒中,插入4个HBV DNA片段而构建成一级检测探针,并结合12个串联重复的二级放大探针和标记的三级探针,建立检测HBV DNA的TRSE液相杂交系统.采用含HBV全基因组的重组质粒DNA对355份肝炎患者或正常人群的血清样本进行检测,评价该系统的效果.结果 在检测探针的一侧插入HBV DNA片段后,建立了HBV DNA的TRSE液相杂交检测系统.检测含HBV全基因组的重组质粒DNA的敏感性为8.33×104拷贝/ml.检测血清样本时,可区分HBeAg阳性与阴性患者间的HBV DNA水平差异.结论 TRSE液相杂交检测HBV DNA系统具有较好的敏感性、特异性和实用性.串联重复核酸序列信号放大的模式具有进一步开发利用的价值。
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