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目的:采用基因芯片技术对严重急性呼吸综合征(SARS)患者进行病毒载量时相监测,为SARS早期诊断提供参考指标。方法:4例确诊非重症SARS患者,在病程的3～30d中,间隔2～6d共7次连续采样。检测标本种类为血液和呼吸道分泌物(痰或鼻、咽拭子)。血标本26份,呼吸道分泌物标本24份,共计50份。对SARS鄄CoV复制酶(replicase)1A、刺蛋白(spikeglycoprotein)和核衣壳蛋白(nucleocapsidprotein)的编码序列区域进行检测和杂交。基因芯片有严格的反应内控体系和防止污染手段。结果:4例SARS患者在病程第4天第1次采样时,2例患儿呼吸道标本阳性,在病程9～18d第2、3次采样时,4例患者的所有标本检测均为强阳性或中等程度阳性。在病程18～23d,第4、5次采样时,所有标本均表现为阳性程度减弱或转为阴性。到病程的22～30d第6、7次采样检测时,4例患者的呼吸道分泌物和血标本已全部无病毒核酸检出。结论:基因芯片技术为SARS早期诊断和病毒载量时相监测提供了有参考意义的实验数据。
OBJECTIVE: To detect the viral load in patients with severe acute respiratory syndrome (SARS) by gene chip technique and to provide a reference for the early diagnosis of SARS. Methods: Four patients with non-severe SARS were enrolled in the study. During the course of 3 ~ 30 days, they were continuously sampled 7 times after 2 ~ 6 days. Specimens were tested for blood and respiratory secretions (sputum or nasal, throat swabs). 26 blood samples, 24 samples of respiratory secretions, a total of 50 copies. The coding sequence regions of SARS-CoV replicase 1A, spikeglycoprotein and nucleocapsid protein were tested and hybridized. Gene chips have a strict reaction control system and prevent pollution means. Results: In the four SARS patients, the first two samples of the course of the first sampling day, two children were positive for respiratory specimens, in the course of 9 ~ 18d the second and third sampling, 4 patients in all specimens were strongly positive or Moderately positive. In the course of 18 ~ 23d, 4,5 samples, all specimens showed a decrease in the degree of positive or turned negative. To the course of the 22 ~ 30d the 6th and 7th sampling test, 4 patients with respiratory secretions and blood samples have been completely virus-free nucleic acid detection. Conclusion: Gene chip technology provides experimental data for early diagnosis of SARS and time-lapse monitoring of viral load.