摘要
目的 为了快速、准确地检测环境中存在的致病菌 ,建立一种采用基因芯片技术对环境中常见致病菌检测和鉴定的实验方法。方法 采用合成后点样的方法把自行设计合成的一系列寡核苷酸探针固定在经过醛基化修饰的显微镜载玻片上 ,制成用于致病菌检测的基因芯片。结果 在相同的条件下 ,扩增了涉及 12个菌属的 15 1株细菌的 16SrDNA基因片段并与基因芯片杂交 ,经Scan Array 30 0 0芯片阅读仪扫描得到特异性的杂交图 ,归纳这些杂交图 ,得到一套属 (种 )特异的典型杂交图谱。待检的样品菌与基因芯片进行杂交 ,得到的杂交结果与典型图谱比对即可判断出样品的种类。用这样的方法对从实际样品中分离的细菌进行检测 ,准确率达到了 96 .2 % (2 5 2 6 )。结论 该项技术的建立为今后更大规模的检测研究奠定了基础 ,可以推广应用于感染性疾病诊断、环境监测、食品卫生监督、商品检验检疫等领域。
Objective To detect the pathogenic bacteria in environment quickly and accurately. Methods A rapid (<3 h) procedure based on the gene chip technology was set up. It composed of three steps: firstly, amplifying a portion of the 16S rDNA to get the target gene; secondly, developing an oligonucleiotide microarray to hybridize with the target genes and to detect the fluorescent signals with ScanArray 3000 scanner. Results One hundred and fifty one strains of bacteria in pure culture belonged to 12 genera that could be successfully discriminated. A series of specific hybridization maps corresponding to each kind of bacterium were obtained. When this procedure was applied to 26 unselected cultures, 25 (96.2%) were correctly identified. Conclusion The accuracy, range, and discriminatory power of the oligonucleotide microarray can be further extended by adding oligonucleotides to the arrays without significantly increasing complexity or cost. [
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2003年第1期74-78,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目 ( 30 0 70 6 43)
86 3资助项目 (SZ0 30 5 )
