摘要
目的 建立快速、灵敏、特异的汉坦病毒基因芯片诊断方法。方法 根据发表的汉坦病毒属 (HV ) 76-118株和R2 2株S基因核苷酸序列 ,设计引物和探针 ,制备寡核苷酸芯片。用Cy3标记核苷和引物 ,运用不对称PCR技术制备单链荧光标记核苷酸片段 ,并与芯片上的寡核苷酸探针杂交 ,荧光扫描仪检测并分析信号。结果 研究制备的基因芯片能够检测汉坦病毒HTN型和SEO型病毒核酸的特异性荧光信号。结论 HV的基因芯片检测具有特异、灵敏、快速的优点。基因芯片的制备和检测技术的建立 ,可为肾综合征出血热等传染病的诊断和预防提供理想的方法。
ObjectiveTo establish a rapid,sensitive and specific dia gnostic assay for Hantavirus with microarray techniques. MethodsSpecific primers and probes were designed accordi ng to the conservative and specific DNA sequence of 76-118 strain and R22 strain .The probes were spotted on glass slides to form microarrays.The Cy3-labled sing le strand DNA fragment prepared by dissymmetical PCR were hybridized with the pr obers on the glass slides.The microarrays were scanned and analyzed with a scann er.ResultsDNA microarray could detect the different typed D NA of HTN and SEO with adequate specificity and sensitivity.ConclusionThe successfully developed DNA microarray and techniques might be a very useful method for diagnosis and prevention,and widely applied in specific pathogen detection of infectious diseases such as hemorrha gic fever with renal syndrome.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2004年第3期268-270,共3页
Chinese Journal of Public Health
基金
全军十五重点课题 ( 0 1L 0 0 6 )
关键词
肾综合征出血热
汉坦病毒
基因芯片
hemorrhagic fever with renal syndrome
hantavir us
DNA microarray
