关键词: 定量PCR;探针杂交;定量试剂

    摘 要:目的 研制一种能用于仪器操作的灵敏、快速、特异的定量试剂. 方法 使用一个生物素标记的上游引物，对HBV DNA及其竞争模板DNA进行不对称PCR扩增，使扩增出的单链带有生物素，并可结合在固定有链亲合素的微孔板的表面.用标有辣根过氧化物酶的待测模板探针和竞争模板探针分别与两个微孔中固定的相应单链PCR产物杂交，而后加底物显色，测定吸光度进行分级定量分析. 结果 该定量方法可以检测10拷贝数的模板，而且对简便处理的标本适应性强;以分级定量的形式定量时，具有良好的重复性. 结论 该试剂适用于仪器操作，可对临床血清中HBV DNA进行分级定量分析.

    A quantitative PCR kit used in automatic genetic amplification instrument

    GUO Yan-Hai YAN Xiao-Jun MENG Xian-Run CUI Da-Xiang HOU Yu HAN Feng-Chan FENG Yong-Qiang ZHANG Ju ZHAO Jin-Rong

    1 Institute of Gene Diagnosis of Chinese PLA，Fourth Military Medical University，Xi'an710033，China，

    2 Xi'an Instrument Factory，Xi'an710082，China

    Keywords:quantitative PCR;probe hybridization;quantita-tive kit

    Abstract:AIM A sensitive，rapid and specific quantitative reagent used in quantitative PCR instrument was researched and the content of HBV DNA in serum was quantified.

    METHODS The HBV DNA and the competitive DNA tem-plate were asymmetrically amplified by a bio-labeled up-stream primer ......