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目的 用合成仪合成60mer寡核苷酸SARS冠状病毒的探针来制作寡核苷酸基因芯片。方法 根据寡核苷酸探针的制作要求,选取SARS冠状病毒的特异序列30条、长度为60mer的片段进行合成,并对合成后的纯化方法与条件进行探索。将经12％变性PAG胶纯化后的探针直接点样制备成12×12阵列的寡核苷酸的基因芯片,初步与处理后的临床样品进行杂交。结果 经PAGE纯化的寡核苷酸探针的纯度高,能满足寡核苷酸芯片制备的要求。临床SABS患者标本进行处理后与寡核苷酸芯片杂交、扫描检测,效果较好。结论 该室合成的60mer的寡核苷酸SARS冠状病毒探针能用于基因芯片的制备与检测,这为该疾病的实验室诊断提供了一种快速、平行的检测方法。
Objective To synthesize an oligonucleotide microarray using a synthetic probe of a 60mer oligonucleotide SARS coronavirus. Methods According to the production of oligonucleotide probes, 30 specific sequences of SARS coronavirus were synthesized and the length of 60mer fragments were synthesized. The purification methods and conditions were explored. After 12% denatured PAG gel purified probes were directly spotted into 12 × 12 arrays of oligonucleotide microarray gene chip, initially with the treatment of clinical samples were hybridized. Results The purity of the PAGE-purified oligonucleotide probes was high enough to meet the requirements of oligonucleotide chip preparation. Clinical SABS patients treated with oligonucleotide chip hybridization, scanning detection, the effect is better. Conclusion The 60 mer oligos SARS coronavirus probe synthesized in this study can be used for gene chip preparation and detection, which provides a rapid and parallel method for the laboratory diagnosis of this disease.