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    • 8. 发明申请
    • ISOLATION AND/OR AMPLIFICATION OF HEPATITIS C VIRUS (HCV) NUCLEIC ACIDS FROM SAMPLES SUSPECTED TO CONTAIN HCV
    • 从包含HCV的样品中分离和/或扩增丙型肝炎病毒(HCV)核酸
    • WO1997037040A2
    • 1997-10-09
    • PCT/NL1997000167
    • 1997-04-03
    • AMSTERDAM SUPPORT DIAGNOSTICS B.V.GOUDSMIT, JaapBELD, Marcellinus, Gualbertus, Hubertus, MariaSOL, Cornelis, Johannes, AndreasBOOM, Willem, René
    • AMSTERDAM SUPPORT DIAGNOSTICS B.V.
    • C12Q01/68
    • C12N15/1006C12Q1/707
    • The invention relates to methods for easily separating single stranded (HCV) nucleic acid material from double stranded nucleic acid material in a sample containing both. By the right choice of at least one chaotropic agent, preferably a guanidine salt, at a selected concentration and other suitable conditions such as chelating agents, pH and the like it is possible to bind double stranded material to a solid phase such as silica particles, whereas single stranded material will not bind under those circumstances. By separating the silica particles from the sample the double stranded nucleic acid material is removed. It can easily be eluted from the silica particles. In a second step the single stranded HCV material may be bound to a solid phase by selecting a different set of conditions. The particles can again be separated from the sample and the single stranded material may now be eluted. For very efficient separations the process may be repeated. Following the separation of the two kinds of nucleic acid, either kind may be amplified. Methods of amplification are provided which do not need sequence data of the material to be amplified. In these methods a primer will be provided with an amplification motif and a random hybridization motif.
    • 本发明涉及在含有两者的样品中容易地从双链核酸材料分离单链(HCV)核酸材料的方法。 通过选择浓度和其它合适条件如螯合剂,pH等的至少一种离液剂,优选胍盐的选择,可以将双链材料结合到固相如二氧化硅颗粒, 而单链材料在这种情况下不会结合。 通过从样品中分离二氧化硅颗粒,除去双链核酸材料。 它可以容易地从二氧化硅颗粒中洗脱出来。 在第二步中,通过选择不同的一组条件,单链HCV材料可以结合固相。 颗粒可以再次与样品分离,单链材料现在可以被洗脱。 为了非常有效的分离,可以重复该过程。 在分离两种核酸后,可以扩增任一种。 提供了不需要放大材料的序列数据的扩增方法。 在这些方法中,引物将具有扩增基序和随机杂交基序。