Grupo de Bioquı́mica de Alimentos, Instituto de Investigacións Mariñas (CSIC), Vigo, Spain.
J Agric Food Chem. 2013 Apr 10;61(14):3488-93. doi: 10.1021/jf400136j. Epub 2013 Mar 25.
Real-time PCR is the most sensitive method for detection and precise quantification of specific DNA sequences, but it is not usually applied as a quantitative method in seafood. In general, benchmark techniques, mainly cycle threshold (Ct), are the routine method for quantitative estimations, but they are not the most precise approaches for a standard assay. In the present work, amplification data from European hake (Merluccius merluccius) DNA samples were accurately modeled by three sigmoid reparametrized equations, where the lag phase parameter (λc) from the Richards equation with four parameters was demonstrated to be the perfect substitute for Ct for PCR quantification. The concentrations of primers and probes were subsequently optimized by means of that selected kinetic parameter. Finally, the linear correlation among DNA concentration and λc was also confirmed.
实时 PCR 是检测和精确定量特定 DNA 序列最敏感的方法，但通常不作为海鲜定量方法应用。一般来说，基准技术，主要是循环阈值（Ct），是定量估计的常规方法，但它们不是标准测定的最精确方法。在本工作中，通过三个 S 型重新参数化方程准确地对欧洲无须鳕（Merluccius merluccius）DNA 样本的扩增数据进行建模，其中四参数 Richards 方程的滞后期参数（λc）被证明是 Ct 的完美替代品，可用于 PCR 定量。随后，通过选择的动力学参数优化引物和探针的浓度。最后，还证实了 DNA 浓度与 λc 之间的线性相关性。
