利用寡核苷酸膜芯片检测SARS病毒

目的：建立寡核苷酸膜芯片技术检测SARS病毒。方法：以SARS冠状病毒RNA聚合酶基因(P基因)作为目的基因．参考WHO公布的特异性引物序列，将SARSRNA进行P基因克隆。以地高辛标记的引物，扩增质粒DNA，在P基因上设计11条寡核苷酸探针，点于尼龙膜，制备膜芯片，与地高辛标记的PCR产物杂交显色。结果：以此特异性引物成功地扩增出了440bp的基因片段；以T载体成功的克隆了此基因；克隆的P基因为靶序列，与膜芯片杂交显示，能与probe1，probe2，probe4，probe6，probe8，probe96条正义链特异性探针稳定杂交，其中Drobe1，probe4，probe9杂交效果最好，作为最后选择的探针。结论：膜芯片技术能快速敏感地鉴定出标本中的SARS病毒RNA。

Detection of SARS virus with oligonucleotide membrane chip

Objective: To set up an oligonucleotide membrane chip technique for detecting SARS virus. Methods: RNA polymerase gene(P gene) being as target gene, SARS RNA P gene was cloned according to the specific primers published by WHO. Then the plasmid DNAs were amplified with digoxin-tagged primers. Eleven specific probes designed in P gene were spotted on the nylon membrane to make a membrane chip which was proceeded hybridization with PCR products. Results: A 440 bp gene fragment was amplified successfully with the primers, and the P gene was cloned with T-vector. The hybridizatin of P gene and probes on the membrane chip showed 6 sense probes(probe1, probe2, probe4, probe6, probe8, probe9) could be hybridized steadily, especially probe1, probe4, probe9 did. Conclusion: The membrane chip can be used to detect SARS virus sensitively and rapidly.