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人免疫缺陷病毒Ⅰ型基因序列分析中的质量控制研究
引用本文:关琪,邢辉,黄海龙,魏民,洪坤学,马鹏飞,梁浩,张卓然,邵一鸣. 人免疫缺陷病毒Ⅰ型基因序列分析中的质量控制研究[J]. 中华检验医学杂志, 2005, 28(3): 322-324
作者姓名:关琪  邢辉  黄海龙  魏民  洪坤学  马鹏飞  梁浩  张卓然  邵一鸣
作者单位:1. 沈阳医学院奉天医院检验科
2. 100050,北京,中国疾病预防控制中心性病艾滋病预防与控制中心病毒免疫室
3. 大连医科大学微生物学教研室
基金项目:国家杰出青年资金项目 ( 39925030 ),国家“十五”科技攻关项目(2001BA705B02)
摘    要: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。

关 键 词:人免疫缺陷病毒 基因序列分析 克隆株 HIV-1 质量控制 样本 DNA序列分析 氨基酸序列 基因进化 核苷酸序列
修稿时间:2004-03-31

Study on quality control in analysis of HIV-1 gene sequences
GUAN Qi,XING Hui,HUANG Hai-long,WEI Min,HONG Kun-xue,MA Peng-fei,LIANG Hao,ZHANG Zhuo-ran,SHAO Yi-ming. Study on quality control in analysis of HIV-1 gene sequences[J]. Chinese Journal of Laboratory Medicine, 2005, 28(3): 322-324
Authors:GUAN Qi  XING Hui  HUANG Hai-long  WEI Min  HONG Kun-xue  MA Peng-fei  LIANG Hao  ZHANG Zhuo-ran  SHAO Yi-ming
Abstract: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.
Keywords:DNA  Viral  HIV long-term survivors  Sequence analysis  Quality control
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