南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (02): 180-183.

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痘苗病毒寡核苷酸检测芯片的设计及研制

王艳1, 马文丽1, 毛向明1, 吴清华1, 李凌1, 王洪敏1, 肖维威1, 郑文岭2   

  1. 1. 第一军医大学分子生物研究所, 广东, 广州, 510515;
    2. 广州军区广州总医院医学实验科, 广东, 广州, 510010
  • 出版日期:2004-02-20 发布日期:2004-02-20
  • 基金资助:
    收稿日期:2003-9-25。
    基金项目:全军十五重大攻关项目
    作者简介:王艳(1978-),女,第一军医大学在读博士研究生,电话:020-61640114-89097

Design and preparation of oligonucleotide microarray for vaccinia virus detection

WANG Yan1, MA Wen-li1, MAO Xiang-ming1, WU Qing-hua1, LI Ling1, WANG Hong-min1, XIAO Wei-wei1, ZHENG Wen-ling2   

  1. 1. 第一军医大学分子生物研究所, 广东, 广州, 510515;
    2. 广州军区广州总医院医学实验科, 广东, 广州, 510010
  • Online:2004-02-20 Published:2004-02-20

摘要: 目的 对痘苗病毒进行寡核苷酸检测芯片的初步研究,为建立寡核苷酸芯片检测该类病毒提供初步研究依据。方法 根据痘苗病毒特异基因设计寡核苷酸探针,人工合成探针后制备寡核苷酸芯片。在病毒感染的不同阶段提取病毒样品DNA及阴性样品DNA,采用限制性显示技术标记,标记样品与芯片杂交后,用Agilent芯片扫描仪检测杂交结果。结果 芯片与病毒样品杂交有较强的杂交信号,而与阴性样品杂交除阳性探针外均无信号。结论 病毒样品与阴性样品杂交信号区别明显,在病毒感染的各个时段也都有明显的杂交信号,反映了寡核苷酸芯片具有较高的特异性和灵敏度。

Abstract: Objective To study the preparation of oligonucleotide microarray for detecting vaccinia virus. Methods Oligonucleotide probes were designed and synthesized according to the specific genes of vaccinia virus. Sample DNA of the virus and the negative control sample were obtained and labeled by restriction display technique, followed by hybridization to the oligonucleotide microarray and scanned by Agilent scanner. Results Strong hybridization signals were detected from the viral DNA hybridized with the microarray, but were absent in the negative sample when positive probes were not used. Conclusion Distinct differences in the hybridization signals between the virus sample and negative sample and between the samples obtained in different phase of infection demonstrate high specificity and sensitivity of the microarray for vaccinia virus detection.

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