用于HIV诊断的Oligo基因芯片研制

目的 研制快速筛查诊断HIV病毒的Oligo芯片。方法以HIV-1B亚型U26942全基因组序列为靶序列,利用DNA Club、Oligo 6.0、BLAST、Alignment等生物信息学软件,设计高度特异、长度均一、具近似熔解温度(T_m)的Oligo 探针,并制备成Oligo芯片。B(U26942)、C(U46016)、F(AF075703)、G(AF061640)四种亚型质粒分别经RD-梯度PCR与随机特异性引物PCR,经荧光标记后与芯片杂交,扫描芯片后进行统计学分析。结果 与结论 获得22条60mer 探针,Oligo芯片特异性、敏感性及稳定性好,为进一步应用提供了条件。

Oligonucleotide microarray for human immunodeficiency virus detection

Objective To develop an oligonucleotide microarray for fast detection of human immunodeficiency virus (HIV). Methods With complete genome sequence of HIV-1 subtype B (U26942) as the target sequence and bioinformatics software such as DNAClub, Oligo6.0, BLAST, Alignment, oligonucleotide probes of high specificity with identical length and similar melting temperature (Tm) were designed and synthesized. Oligonucleotide microarray was prepared using Cartesian Microarrayer. Using the plasmids of HIV-1 subtype B(U26942), C(U46016), F(AF075703), G(AF061640) and restriction display technique, Cy3-labeled HIV DNA fragments were amplified and the hybridization results were scanned and analyzed with Array-Pro. Results and Conclusion Twenty-two optimized oligonucleotide microarray probes were obtained and used to prepare the oligonucleotide microarray for further screening studies. The microarray prepared significantly enhanced the sensitivity, reliability and speed of DNA assay, and possesses the potential for application in clinical setting.