SARS冠状病毒基因芯片的制作与初步临床样本验证

Fabrication and application of SARS coronavirus gene chips for preliminary clinical sample testing

为了实现对非典(SARS)冠状病毒感染患者的早期诊断,构建了一套以基因芯片为基础的SARS冠状病毒基因分析检测系统。首先从患者痰、血样本中快速分离出SARS冠状病毒的RNA,然后通过巢式RT-PCR对分离的RNA进行扩增获得特定的DNA片段,最后让固定在基因芯片上的DNA探针与扩增产物进行杂交以检验扩增产物的序列匹配性,达到最终确认SARS冠状病毒感染患者的目的。对首期两例患者痰样本和两例全血样本的分析结果表明,这4例患者样本中均存在SARS冠状病毒。

A gene chipbased analytical system was developed for potential early detection of severe acute respiratory syndrome (SARS) coronavirus presented in the collected clinical samples. Initially, SARS coronavirus RNAs were extracted from both sputum and whole blood. The cDNAs were then produced through reversetranscription process followed by nested PCR to generate sufficient amount of amplicons with expected sizes. Finally, those amplicons were allowed to hybridize against the SARS coronavirusspecific DNA probes immobilized on the glass substrate for confirming the presence of the SARS coronavirus in the collected clinical samples. Four clinical samples (two sputum and two whole blood) have been tested with the gene chips and the results indicate that the SARS coronavirus has been presented in all four samples.