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        Development and Evaluation of a Duplex Real-Time PCR Assay With a Novel Internal Standard for Precise Quantification of Plasma DNA

        Dan Chen,Shi-yang Pan,Erfu Xie,Li Gao,Huaguo Xu,Wenying Xia,Ting Xu,Peijun Huang 대한진단검사의학회 2017 Annals of Laboratory Medicine Vol.37 No.1

        Background: Circulating levels of cell-free DNA increase in many pathologic conditions. However, notable discrepancies in the quantitative analysis of cell-free DNA from a large number of laboratories have become a considerable pitfall, hampering its clinical application. Methods: We designed a novel recombinant DNA fragment that could be applied as an internal standard in a newly developed and validated duplex real-time PCR assay for the quantitative analysis of total cell-free plasma DNA, which was tested in 5,442 healthy adults and 200 trauma patients. Results: Compared with two traditional methods, this novel assay showed a lower detection limit of 0.1 ng/mL, lower intra- and inter-assay CVs, and higher accuracy in the recovery test. The median plasma DNA concentration of healthy males (20.3 ng/mL, n=3,092) was significantly higher than that of healthy females (16.1 ng/mL, n=2,350) (Mann-Whitney two-sample rank sum test, P<0.0001). The reference intervals of plasma DNA concentration were 0-45.8 ng/mL and 0-52.5 ng/mL for healthy females and males, respectively. The plasma DNA concentrations of the majority of trauma patients (96%) were higher than the upper normal cutoff values and were closely related to the corresponding injury severity scores (R2=0.916, P<0.0001). Conclusions: This duplex real-time PCR assay with a new internal standard could eliminate variation and allow for more sensitive, repeatable, accurate, and stable quantitative measurements of plasma DNA, showing promising application in clinical diagnosis.

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        Separable Recursive Gradient Algorithm for Dynamical Systems Based on the Impulse Response Signals

        Ling Xu,Feng Ding,Erfu Yang 제어·로봇·시스템학회 2020 International Journal of Control, Automation, and Vol.18 No.12

        The identification for process control systems is considered in this paper based on the impulse response signals from the discrete measurements. By taking advantage of impulse signals and through the model parameter decomposition, two dependent identification models are constructed and two identification sub-algorithms are presented based on the nonlinear gradient optimization. In terms of the associated items of the parameters to be estimated between two derived sub-algorithms, a separable recursive gradient parameter estimation method is proposed by designing an interactive and recursive estimation. The performance tests and the comparison experiments are carried out by the simulation examples.

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