基因芯片用于SARS早期诊断和病毒载量时相监测研究  

Early diagnosis and monitoring the viral load phases in SARS patients by gene chip technology

作　　者：肖白[1] 王辰[1] 吕月平[1] 谭淑珍[1] 闫梅[1] 丁洁[1] 王臻[1] 刘敬忠[1] 周一鸣[2,3] 王栋[2,3] 高华方[2,3] 陶生策[2,3] 杜建宇[4] 李泽[4] 蒋迪[4] 张琼[4] 范肖冬[4] 张川[4] 程京[2,3,4] 任丽丽[5] 杨仁全[5] 洪涛[5] 王健伟[5] 

机构地区：[1]国家SARS防治紧急科技行动北京组:首都医科大学附属北京朝阳医院 [2]清华大学生物科学与技术系,生物膜与膜生物工程国家重点实验室 [3]生物芯片北京国家工程研究中心,北京100084 [4]北京博奥生物芯片有限责任公司 [5]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《诊断学理论与实践》2004年第3期200-202,205,共4页Journal of Diagnostics Concepts & Practice

基　　金：国家SARS防治紧急科技行动基金及国家SARS科技攻关项目(编号:2003AA208102);国家教育振兴计划项目;清华大学百名人才引进计划项目及科技部功能基因组与生物芯片重大专项资金资助。

摘　　要：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 provide reference index for early diagnosis of SARS and to monitor the phases of viral load in SARS patients by gene chip technology. Methods Four patients with non-serious SARS were followed up for 30 days. The samples of blood and respiratory tract secretion were collected every 2-6 days for seven times, including 26 EDTA anticoagulated peripheral blood samples and 24 respiratory tract secretion (sputum or nasopharyngeal swab or pharyngeal swab) samples. Four sets of the RT-nested PCR primers and the gene chip probes were designed in SARS-CoV RNA region of coding replicase 1A, spike glycoprotein and nucleocapsid protein. The dUTP/UNG was added to the amplification system for avoiding the contamination. The negative control, the positive control, the blank control and the system internal control were strictly designed in the gene chip. Results SARS-CoV RNA was detected only in the respiratory tract samples collected at day 4 after the onset of symptoms in 2 of 4 patients , the peak viral load with strong or medium positive expressions at day 9-18 was showed in all the samples obtained from the 4 patients and the load dropped at day 18-23 and dispersed at day 22-30 in total samples. Conclusions SARS-CoV detection by gene chip technology may be a use ful reference for early diagnosis and monitoring of the viral load in SARS patients.

关 键 词：严重急性呼吸综合征 SARS病毒 基因芯片 病毒载量时相监测 诊断 RT-PCR 

分 类 号：R563.1[医药卫生—呼吸系统]