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1. 发明申请

WO2004042085A3 METHOD FOR QUANTIFYING A RATIO BETWEEN AT LEAST TWO NUCLEIC ACID SEQUENCES 审中-公开
标题翻译：用于定量至少两个核酸序列之间的比例的方法
公开(公告)号：WO2004042085A3
公开(公告)日：2004-08-05
申请号：PCT/NL0300780
申请日：2003-11-07
申请人： PRIMAGEN HOLDING BV , TIMMERMANS EVELINE CATHERINA A , VAN GEMEN BOB
发明人： TIMMERMANS EVELINE CATHERINA A , VAN GEMEN BOB
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6851 , C12Q2527/113
摘要： 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.
摘要翻译：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感兴趣的核酸的量的初始比例的方法，包括： - 扩增所述扩增反应中的所述目的核酸; - 在所述反应中在至少两个不同时间点测量至少两种感兴趣的核酸的量; - 从所述测量中的至少两个确定所述至少两个感兴趣的核酸的扩增速率; - 将比率与参考值进行比较; 以及 - 从所述比较确定所述样品中所述至少两种感兴趣的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感兴趣的核酸的扩增和/或检测限之前达到感兴趣的所有核酸的可检测水平。利用本发明的方法，可以确定比率之间的小差异。此外，可以确定初始核酸比的直接测量。本发明的方法例如适用于确定细胞生物的功能，用于确定疾病的分期，以及用于确定药物或候选化合物的治疗活性和/或可能的副作用。

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