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[一种利用扩增片段的质谱分析进行人线粒体DNA法医分型的新方法:PLEX-ID自动遗传分析系统]

[A new approach to forensic medical typing of human mitochondrial DNA with the use of mass-spectrometric analysis of amplified fragments: PLEX-ID automated genetic analysis system].

作者信息

Ivanov P L

出版信息

Sud Med Ekspert. 2010 May-Jun;53(3):46-51.

PMID:20734792
Abstract

A new approach to typing human mitochondrial DNA (mtDNA) is described using multiplex PCR in combination with genetic analysis assisted by high performance mass-spectrometry based on an automated PLEX-ID ESI-TOF analyzer (Abbot Molecular, USA). The proposed test-system is designed for DNA analysis in a standard 96-well plate and allows to simultaneously treat up to 12 samples. All stages of the analysis are fully automated, from preparation and purification of amplicons to quality control and interpretation of final results. The in-built software makes possible comprehensive analysis of the data obtained including decoding of primary results; search for, comparison and registration of polymorphism profiles in relevant databases, both directly in the nucleotide composition format and indirectly in the form of nucleotide sequences. The entire process from sample preparation to final report takes less than 8 hours to be completed. It is concluded that the PLEX-ID analytical system may be used to resolve complex problems due to the possibility to combine gene typing and individualization of the biological sample in the framework of a single analytical process starting from the sampling procedure.

摘要

本文描述了一种利用多重聚合酶链式反应(PCR)结合基于美国雅培分子公司的自动化PLEX-ID电喷雾电离飞行时间(ESI-TOF)分析仪的高性能质谱辅助基因分析来对人类线粒体DNA(mtDNA)进行分型的新方法。所提出的检测系统专为在标准96孔板中进行DNA分析而设计,可同时处理多达12个样本。从扩增子的制备和纯化到最终结果的质量控制和解读,分析的所有阶段均完全自动化。内置软件使得对所获数据进行全面分析成为可能,包括对初步结果的解码;在相关数据库中直接以核苷酸组成格式以及间接以核苷酸序列形式搜索、比较和记录多态性图谱。从样本制备到最终报告的整个过程不到8小时即可完成。得出的结论是,由于能够在从采样程序开始的单个分析过程框架内将基因分型与生物样本个体化相结合,PLEX-ID分析系统可用于解决复杂问题。

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