军事医学2017，Vol.41Issue(2)：130-134,5.DOI:10.7644/j.issn.1674-9960.2017.02.012
腺病毒分型基因芯片方法的建立和验证
Dvelopment of a DNA-based microarray for detection of human adenovirus serotypes
摘要
Abstract
Objective To develop a chemiluminescence imaging DNA microarray method for simultaneous,quick and accurate detection of serotypes of human adenovirus (HAdV ),namely,HAdV3,HAdV7,HAdV11,HAdV14 and HAdV55.Methods Based on the specific gene sequences in the conserved region of adenovirus from GenBank, oligonucleotide primers and probes were designed and synthesized to prepare the oligonucleotide microarray.The specific genomic sequences were amplified by multiplex PCR method.The multiplex PCR amplification products were hybridized with the specific probes of microarray that was scanned after washing and chemiluminescenceb before the result was analyzed.After optimization of the multiple PCR system,hybridization reactions and conditions of chemiluminescence,the specificity,sensitivity and reproducibility of the chip were evaluated.Results The microarray displayed high sensitivity, specificity and reproducibility.The minimum detection limit of plasmidg DNA was 3 ×103 copies/reaction.The microarray detection results of 38 clinical samples were approximately consistent with those using the direct sequencing method(37 /38).Conclusion A chemiluminescence imaging DNA microarray method for quick,sensitive and specific detection of five serotypes of HAdV is established,which can provide a new means for detecting serotypes of HAdV.关键词
腺病毒科/分型/基因芯片/化学发光Key words
adenoviridae/serotyping/gene chip/chemiluminescent分类
医药卫生引用本文复制引用
陈晓飞,刘琪琦,周伟,张五星,王升启..腺病毒分型基因芯片方法的建立和验证[J].军事医学,2017,41(2):130-134,5.基金项目
国家科技重大专项"重大新药创制"资助项目(2015ZX09J15105-001-001) （2015ZX09J15105-001-001）
