人免疫缺陷病毒Ⅰ型基因序列分析中的质量控制研究

Study on quality control in analysis of HIV-1 gene sequences

目的 研究DNA序列分析中各种影响因素的作用,建立排除污染、进行序列质量控制的方法。方法 通过对 950份HIV 1样品DNA基因序列结果的分析,查找序列读取及序列分析中存在的各种影响因素,对各种可能导致污染的原因进行分析和解释。结果 在使用各种软件进行序列分析时,两样本之间的基因距离为 0;两样本所测区段的核苷酸或氨基酸序列完全一致或相差甚微;样本与实验室内所构建的克隆株之间的基因距离过近,同源性达到 99%以上;两个独立传播的群体之间个别样本的互混等指标均提示存在污染的可能。结论 构建基因进化树和将样本的核苷酸序列翻译成蛋白质的氨基酸序列后构建共享序列,是一种很好的发现序列质量问题、进行序列质量控制的方法。

Objective To study various factors influencing DNA sequencing and sequence analysis, and explore the method of avoiding contamination and obtaining sequence quality assurance Methods Through sequencing 950 HIV 1 samples and carefully analyzing the data, we try to identify various factors influencing DNA sequencing and sequence analysis, and to analyze the potential causes of sequence contamination Results When sequences were phylogeneticlly analyzed by software, some results, for example, the gene distance between two samples was 0, the amino acid sequences or nucleotide sequences of two samples were identical, the gene distance between a sample and the developed clone in the same laboratory was too close, or some sequences from unrelated people were in confusion in phylogenetic tree, might suggest the possibility providing some clues to find sequence contamination of sequence contamination Conclusions Phylogenetic analysis of nucleotide sequences will provide some clues to find sequence contamination.