转基因成分定量检测数据分析及不确定度评估现状概述

摘要

在利用实时荧光定量(real-time fluorescence quantitative polymerase chain reaction, qRT-PCR)检测转基因成分的含量时,数据分析的规范化与标准化对保证实验数据的准确性及实验室间数据的可比性具有重要意义。基于目前很多国家和地区对转基因产品有强制性标识要求的现状,测量不确定度在进出口食品的转基因检验中作用显著。本文在分析 qRT-PCR 实验至少应提供的信息(the minimum information for publication of quantitative Real-time PCR experiments(MIQE))数据分析的基础上,综合阐述了qRT-PCR转基因成分定量数据分析中众多因素对定量准确性的影响,以及 qRT-PCR 转基因成分定量不确定度来源主--素及其相应的评估策略。尽管 MIQE为 qRT-PCR实验设计及数据处理提供了参考,但目前仍然有许多实验设计甚至已经发表的论文并未完全遵照相关要求,从而导致实验结果偏差甚至错误,其数据处理的某些环节尚缺乏一致性的要求。由于对一些影响因子缺少定量描述,在转基因成分定量检测的不确定度分析方面还缺乏一个通行的方法。%ABSTRACT:During the quantitative real-time PCR experiments for dection of genomic modified (GM) ingredients, the normalization of data processing is important for the accuracy of experimental results and comparability among different laboratories. Since it is now required to label the GM products in many countries and regions, the uncertainty of measurement is vital for the GM detection in the imports and exports. Based on the minimum information for publication of quantitative real- time PCR experiments(MIQE), the affects of various factors for quantitative accuracy, as well as the main resources of measurement uncertainty and corresponding evaluations were elucidated. Although MIQE provides reference for qRT-PCR experimental design and data processing, but at present many experimental design and published papers still did not fully comply with the relevant requirements, which lead to the result deviation even error. Some links of data processing are short in consistency requirements. Due to the lack of quantitative description for some factors, a generally accepted method of uncertainty analysis on genetically modified ingredients quantitation has not been developed.