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高通量二代测序技术在病毒性肝癌HBV-DNA与宿主基因整合位点研究中的应用 被引量:1
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作者 麦庆锋 张润玲 +4 位作者 曾君 吴梓坤 程欢 吴英松 吴意 《热带医学杂志》 CAS 2022年第2期154-159,183,共7页
目的运用高通量二代测序技术(NGS)捕获病毒性肝癌组织中乙型肝炎病毒(HBV)DNA与宿主基因整合位点,为临床疾病诊断治疗提供可靠的依据。方法采用高通量二代测序技术对5例原发性病毒性肝癌组织DNA进行全基因组测序,使用Burrows-Wheeler Al... 目的运用高通量二代测序技术(NGS)捕获病毒性肝癌组织中乙型肝炎病毒(HBV)DNA与宿主基因整合位点,为临床疾病诊断治疗提供可靠的依据。方法采用高通量二代测序技术对5例原发性病毒性肝癌组织DNA进行全基因组测序,使用Burrows-Wheeler Aligner(BWA)和Picard-tools生物信息学软件,采用BWA-MEM比对方法进行数据处理和分析,从宿主基因组DNA序列中筛选出HBV基因的整合位点。结果5个样品共发现3513454个单核苷酸多态性(SNP)、838846个插入缺失标记(InDel)、平均1965个结构变异(SV)和3639个拷贝数变异(CNV),其中SNP 99.48%出现在dbSNP数据库里,95.26%在千人基因组计划的数据库中,新发现的SNP有17768个;InDel 93.80%出现在dbSNP数据库里,54.23%在千人基因组计划的数据库中,新发现的InDel有51307个;SV平均包括0个插入变异、690个缺失变异、257个倒位变异、762个染色体内易位变异和255个染色体间易位变异。结论HBV-DNA与宿主基因整合位点具有随机性,且插入的核苷酸片断大小不一,导致宿主肝细胞基因组发生突变、缺失和插入等结构变异而产生致癌作用。 展开更多
关键词 高通量 二代测序技术 乙型肝炎病毒 肝癌 基因整合位点
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Comparison of effectiveness of whole viral,N and N199 proteins by ELISA for the rapid diagnosis of severe acute respiratory syndrome coronavirus 被引量:2
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作者 GUO Zhong-min LU Jia-hai +8 位作者 HAN Wen-yu LIU Ze-yu LI Guo-wei LIAO Jia-wei WANG Shu-min wu ying-song ZHENG Huan-ying ZHONG Nan-shan ZHU Xing-quan 《Chinese Medical Journal》 SCIE CAS CSCD 2007年第24期2195-2199,共5页
Background Although severe acute respiratory syndrome (SARS) has been controlled, the subsequently emerging sporadic cases in 2004 emphasize the necessity of developing a rapid diagnostic method, which would be of g... Background Although severe acute respiratory syndrome (SARS) has been controlled, the subsequently emerging sporadic cases in 2004 emphasize the necessity of developing a rapid diagnostic method, which would be of great help in clinical diagosis and also wild host screening. This study aims to establish an effective and rapid serological tool for the diagnosis of SARS-CoV by comparison among whole viral, N and N199 proteins by ELISA. Methods SARS-CoV N and N199 (a truncated nucleocapsid gene) genes were cloned, expressed, identified by Western blotting, and applied in screening of human and swine samples. Sera of SARS convalescent-phase patients, normal human sera, sera of patients with other respiratory diseases, and swine sera were screened by ELISA, with whole SARS-CoV F69, N and N199 proteins as antigens. Results The sensitivity and specificity of N and N199 proteins in human sera diagnosis were approximate (P=-0.743), which was higher than whole viral protein but the difference was not significant (P=-0.234). The N199 protein proved to be more specific in swine sera screening than whole viral and N protein (P〈0.001). Conclusion N199 protein is feasible in both clinical diagnosis and SARS-CoV reservoir screening. 展开更多
关键词 severe acute respiratory syndrome virus N199 enzyme-linked immunosorbent assay
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可溶性fms样酪氨酸激酶1时间分辨荧光免疫分析法的建立
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作者 吴鸿峰 邓昊 +5 位作者 翁孔燕 黄颖而 陈佳灵 李志雄 吴英松 郝文波 《现代免疫学》 CAS 北大核心 2022年第5期375-380,共6页
为建立基于磁性微球可溶性fms样酪氨酸激酶1(soluble fms-like tyrosine kinase 1,sFlt-1)的时间分辨荧光免疫分析法(sFlt-1 time-resolved fluoroimmunoassay, sFlt-1-TRFIA),用磁珠偶联抗sFlt-1单克隆抗体,Eu^(3+)(铕)标记抗sFlt-1单... 为建立基于磁性微球可溶性fms样酪氨酸激酶1(soluble fms-like tyrosine kinase 1,sFlt-1)的时间分辨荧光免疫分析法(sFlt-1 time-resolved fluoroimmunoassay, sFlt-1-TRFIA),用磁珠偶联抗sFlt-1单克隆抗体,Eu^(3+)(铕)标记抗sFlt-1单克隆抗体,采用双抗体夹心法建立基于磁珠的sFlt-1-TRFIA。结果显示,自制试剂的检测灵敏度为10.78 pg/mL,线性范围10.78~25 000 pg/mL;样品的回收率为98.23%~107.71%,分析内变异系数(intraassay coefficient of variation, CV)为5.29%~6.50%,分析间变异系数(intreassay coefficient of variation, CV)为6.55%~8.23%;与电化学发光法的检测结果比较,相关系数r^(2)值为0.971 9。由此,基于磁性微球建立的sFlt-1-TRFIA灵敏度、特异度以及准确度等均符合临床应用要求,有望用于临床血清样本中sFlt-1浓度的检测。 展开更多
关键词 磁性微球 可溶性fms样酪氨酸激酶1 时间分辨荧光免疫分析法 先兆子痫
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