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首页> 外文期刊>Genomics >Identification of pathogenic Helicobacter species by chaperonin-60 differentiation on plastic DNA arrays.
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Identification of pathogenic Helicobacter species by chaperonin-60 differentiation on plastic DNA arrays.

机译：通过分子伴侣60在塑料DNA阵列上的分化鉴定致病性幽门螺杆菌。

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摘要

A microarray method for bacterial species identification based on cpn60 and 16S rDNA hybridization was developed. Specific cpn60 or 16S rDNA oligonucleotides from various Helicobacter or Campylobacter species were printed and immobilized onto a proprietary plastic solid support. Using universal primers, fragments derived from either cpn60 or 16S rDNA genes from single isolates or from a complex human waste sludge DNA sample spiked with Helicobacter pylori were biotinylated and hybridized to the plastic slide. Subsequent querying with a streptavidin-horseradish peroxidase conjugate followed by color development using tetramethylbenzidine resulted in accurate Helicobacter species identification with no cross-hybridization to either the 16S rDNA or the cpn60 sequence of a closely related strain of Campylobacter jejuni. The combination of a nonfluorescence visual detection system with a polymer-based DNA microarray slide has resulted in a molecular tool that should prove useful in numerous applications requiring rapid, low-cost bacterial species identification.
机译：建立了基于cpn60和16S rDNA杂交技术的细菌种鉴定微阵列方法。将来自各种幽门螺杆菌或弯曲杆菌属的特定cpn60或16S rDNA寡核苷酸印刷并固定在专有的塑料固体支持物上。使用通用引物，将源自单个分离株的cpn60或16S rDNA基因片段或掺入幽门螺杆菌的复杂人类废物污泥DNA样品的片段进行生物素化处理，并与塑料载玻片杂交。随后用链霉亲和素-辣根过氧化物酶偶联物进行查询，然后使用四甲基联苯胺显色，可准确鉴定幽门螺杆菌，而不会与空肠弯曲杆菌紧密相关菌株的16S rDNA或cpn60序列交叉杂交。非荧光视觉检测系统与基于聚合物的DNA微阵列载玻片的组合，已经产生了一种分子工具，该工具应证明可用于需要快速，低成本细菌种类鉴定的众多应用中。

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