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    • 5. 发明申请
    • METHOD FOR QUANTIFYING A RATIO BETWEEN AT LEAST TWO NUCLEIC ACID SEQUENCES
    • 量化至少两个核酸序列之间比率的方法
    • WO2004042085A2
    • 2004-05-21
    • PCT/NL2003/000780
    • 2003-11-07
    • PRIMAGEN HOLDING B.V.TIMMERMANS, Eveline, Catherina, Anna, ClasinaVAN GEMEN, Bob
    • TIMMERMANS, Eveline, Catherina, Anna, ClasinaVAN GEMEN, Bob
    • C12Q1/68
    • C12Q1/6851C12Q2527/113
    • The invention provides a method for quantifying an initial ratio of the amounts of at least two nucleic acids of interest in a sample by means of a multiplex nucleic acid amplification reaction, comprising:- amplifying said nucleic acids of interest in said amplification reaction; - measuring the amount of at least two nucleic acids of interest at at least two different time points in said reaction; - determining from at least two of said measurements the amplification rate of said at least two nucleic acids of interest; - comparing said rates with a reference; and - determining from said comparison said initial ratio of the amounts of said at least two nucleic acids of interest in said sample. Preferably, at least one variable factor in said nucleic acid amplification reaction is adjusted in order to allow detectable levels of all nucleic acids of interest to be reached before an amplification and/or detection limit of one or more of the nucleic acids of interest is reached. With a method of the invention, small differences between ratios can be determined. Moreover, direct measurement of the initial nucleic acid ratio can be determined. A method of the invention is for instance suitable for determining functioning of a cellular organism, for determining the staging of a disease, and for determining therapeutic activity and/or possible side-effects of a medicament or candidate compound.
    • 本发明提供了一种通过多重核酸扩增反应来定量样品中至少两种目的核酸的量的初始比例的方法,其包括: - 扩增 对所述扩增反应的兴趣; - 在所述反应中的至少两个不同时间点测量至少两种感兴趣核酸的量; - 从至少两次所述测量中确定所述至少两种目标核酸的扩增速率; - 将所述比率与参考值进行比较; 和 - 根据所述比较确定所述样品中所述至少两种感兴趣核酸的量的初始比例。 优选地,调节所述核酸扩增反应中的至少一个可变因子,以便在达到感兴趣的一种或多种核酸的扩增和/或检测极限之前达到可检测水平的所关注的所有核酸 。 利用本发明的方法,可以确定比率之间的微小差异。 此外,可以确定初始核酸比率的直接测量。 本发明的方法例如适用于确定细胞生物体的功能,确定疾病的分期以及确定药物或候选化合物的治疗活性和/或可能的副作用。
    • 6. 发明申请
    • STORING AND DETECTING NUCLEIC ACID ADMINISTERED TO A SOLID CARRIER
    • 储存和检测核酸的固体载体
    • WO2003080869A2
    • 2003-10-02
    • PCT/NL2003/000491
    • 2003-07-03
    • PRIMAGEN HOLDING B.V.DE ROOIJ, Esther, ReginaDE BAAR, Marinus, Petrus
    • DE ROOIJ, Esther, ReginaDE BAAR, Marinus, Petrus
    • C12Q1/68
    • C12Q1/6806C12Q1/703C12Q2545/114C12Q2565/518
    • The invention provides a method for detecting a nucleic acid of interest in at least one sample, comprising administering said sample to a solid carrier capable of at least in part absorbing said sample, drying said carrier, providing at least a representative part of said carrier to a nucleic acid isolation solution so that a representative amount of said nucleic acid is extracted from said carrier, and detecting said representative amount of said nucleic acid. With a method of the invention a sample, such as a body fluid sample, is stabilized in such a way that it can be shipped from the site of taking (for instance local hospital or lab in a less-developed country) and be sent to a service testing laboratory elsewhere in the world by normal logistics means. Preferably at least 100 µl , more preferably at least 250 µl of sample is administered to said carrier, in order to detect low titers of nucleic acid of interest. A kit of parts for detecting, identifying and/or quantifying a nucleic acid of interest in a sample, comprising a solid carrier capable of at least in part absorbing said sample, and a nucleic acid isolation solution, is also herewith provided.
    • 本发明提供了用于检测至少一种样品中感兴趣的核酸的方法,包括将所述样品施用至能够至少部分吸收所述样品的固体载体,干燥所述载体,提供在 将所述载体的至少一个代表性部分与核酸分离溶液接触,从而从所述载体中提取代表性量的所述核酸,并检测所述代表性量的所述核酸。 采用本发明的方法,诸如体液样本的样本稳定在可从服用部位(例如当地医院或欠发达国家的实验室)运送并被发送到 通过正常的物流手段在世界其他地方的服务测试实验室。 优选地,将至少100微升,更优选至少250微升的样品施用于所述载体,以检测目标核酸的低滴度。 还提供了用于检测,鉴定和/或定量样品中感兴趣的核酸的部分试剂盒,其包含能够至少部分吸收所述样品的固体载体和核酸分离溶液。