Weber D G, Sahm K, Polen T, Wendisch V F, Antranikian G
Institute of Technical Microbiology, Hamburg University of Technology, Hamburg, Germany.
J Appl Microbiol. 2008 Oct;105(4):951-62. doi: 10.1111/j.1365-2672.2008.03799.x. Epub 2008 Sep 10.
The design and evaluation of an oligonucleotide microarray in order to detect and identify viable bacterial species that play a significant role in beer spoilage. These belong to the species of the genera Lactobacillus, Megasphaera, Pediococcus and Pectinatus.
Oligonucleotide probes specific to beer spoilage bacteria were designed. In order to detect viable bacteria, the probes were designed to target the intergenic spacer regions (ISR) between 16S and 23S rRNA. Prior to hybridization the ISR were amplified by combining reverse transcriptase and polymerase chain reactions using a designed consenus primer. The developed oligonucleotide microarrays allows the detection of viable beer spoilage bacteria.
This method allows the detection and discrimination of single bacterial species in a sample containing complex microbial community. Furthermore, microarrays using oligonucleotide probes targeting the ISR allow the distinction between viable bacteria with the potential to grow and non growing bacteria.
The results demonstrate the feasibility of oligonucleotide microarrays as a contamination control in food industry for the detection and identification of spoilage micro-organisms within a mixed population.
设计并评估一种寡核苷酸微阵列,用于检测和鉴定在啤酒变质过程中起重要作用的活细菌种类。这些细菌属于乳杆菌属、巨球形菌属、片球菌属和果胶杆菌属。
设计了针对啤酒变质细菌的寡核苷酸探针。为了检测活细菌,探针设计为靶向16S和23S rRNA之间的基因间隔区(ISR)。在杂交之前,使用设计的共有引物通过逆转录酶和聚合酶链反应相结合来扩增ISR。所开发的寡核苷酸微阵列能够检测活的啤酒变质细菌。
该方法能够在含有复杂微生物群落的样本中检测和区分单个细菌种类。此外,使用靶向ISR的寡核苷酸探针的微阵列能够区分有生长潜力的活细菌和无生长能力的细菌。
结果证明了寡核苷酸微阵列作为食品工业中污染控制手段用于检测和鉴定混合群体中腐败微生物的可行性。