不同品牌核酸提取仪在新冠病毒核酸检测中的差异性分析

To analyze the difference in novel Coronavirus nucleic acid extraction in different brands extraction apparatus

目的比较不同品牌核酸提取仪所提核酸质量以及分析不同品牌扩增仪扩增结果之间的差异。方法选用6种品牌核酸提取仪和1种手工提取试剂对20例健康人群咽拭子样本进行核酸提取,并在荧光定量PCR仪上进行扩增,综合比较各组样本核酸浓度、纯度和Ct值。后将A品牌提取仪所提核酸在5个不同品牌荧光定量PCR仪上进行扩增,比较同一组样本在不同扩增仪之间的差异。结果提取核酸浓度从高到低依次为:A>C>E>D>F>B>G,各品牌之间比较差异具有统计学意义(P<0.05);提取核酸纯度从大到小依次为:C>E>F>B>G>A>D,各品牌之间比较差异具有统计学意义(P<0.05),B、E、F、G四种品牌纯度测定值在1.8~2.0范围内;提取核酸扩增Ct值从小到大依次为:A<C<B<F<G<D<E,F=4.325,各品牌之间比较差异具有统计学意义(P<0.05);同一核酸不同PCR仪上进行扩增结果Ct值从小到大依次为:Ⅳ<Ⅴ<Ⅰ<Ⅲ<Ⅱ,各品牌之间比较差异具有统计学意义(P<0.05)。最后使用不同提取仪对第三方阳性质控品进行提取并进行扩增,G品牌所提质控品Ct值在7个品牌中最小,B品牌质控品Ct值在7个品牌中最大。结论不同品牌核酸提取仪所提核酸均能满足荧光定量RT-PCR的要求,但各品牌之间提取质量和扩增效率仍存在一定的差别,实验室选用之前需要做好评价工作,充分了解使用仪器的性能。

Objective To compare the nucleic acid qualities in different brands extraction apparatus and analyze the Ct values difference in differnent fluorescence quantitative PCR instuments.Methods Nucleic acid were extracted from 20 healthy people's Throat swab samples by 6 kinds of brands nucleic acid extraction apparatus and 1 kinds of brand manual extraction reagent,and amplified by fluorescence quantitative PCR instrument,then the concentration and purity of nucleic acid and Ct values of samples in each group were compared.Besides,the nucleic acid which extracted from brand A extractor was amplified on 5 kinds of brands fluorescence quantitative PCR instruments and the Ct values differences were compared.Results The extracted nucleic acid concentrations were ranked as:A>C>E>D>F>B>G,F=13.19,P<0.0001,the difference were statistically significant;The purity of nucleic acid were ranked as:C>E>F>B>G>A>D,F=41.84,P<0.0001,the difference were statistically significant,B,E,F and G are in the range of 1.8-2.0;The Ct values of nucleic acid amplification were ranked as:A<C<B<F<G<D<E,F=4.325,P=0.0005,the difference were statistically significant;The Ct value of different PCR instruments of the same nucleic acid were ranked as:Ⅳ<Ⅴ<Ⅰ<Ⅲ<Ⅱ,F=30.26,P<0.0001.Finally,different extractors for positive quality control products which from the third party were extracted and amplified.The Ct value of G brand was the smallest and B brand was the largest.Conlusions Different brands of nucleic acid extractors can meet the requirements of fluorescence quantitative RT-PCR,but there are some differences in extraction quality and amplification efficiency among brands.So it is necessary for each laboratory to do a evaluation before used to fully understand the performance of the instrument.