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首页> 外文期刊>Nucleic Acids Research >Quenched autoligation probes allow discrimination of live bacterial species by single nucleotide differences in rRNA.
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Quenched autoligation probes allow discrimination of live bacterial species by single nucleotide differences in rRNA.

机译：淬灭的自连接探针可以通过rRNA中的单核苷酸差异来区分活细菌。

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摘要

Quenched autoligation (QUAL) probes are a class of self-reacting nucleic acid probes that give strong fluorescence signal in the presence of fully complementary RNAs and selectivity against single nucleotide differences in solution. Here, we describe experiments designed to test whether QUAL probes can discriminate between bacterial species by the detection of small differences in their 16S rRNA sequences. Probes were introduced into live cells using small amounts of detergent, thus eliminating the need for fixation, and fluorescence signal was monitored both by microscopy and by flow cytometry without any washing steps. The effects of probe length, modified backbone, probe concentration and growth state of the bacteria were investigated. The data demonstrate specific fluorescence discrimination between three closely related bacteria, Escherichia coli, Salmonella enterica and Pseudomonas putida, based on single nucleotide differences in their 16S rRNA. Discrimination was possible with cells in mid-log phase or in lag phase. These results suggest that QUAL probes may be useful for rapid identification of microorganisms in laboratory and clinical settings.
机译：淬灭自连接（QUAL）探针是一类自反应核酸探针，在存在完全互补的RNA时会发出强烈的荧光信号，并且对溶液中的单核苷酸差异具有选择性。在这里，我们描述了旨在测试QUAL探针是否能够通过检测其16S rRNA序列中的细微差异来区分细菌物种的实验。使用少量去污剂将探针引入活细胞，从而无需固定，并且无需任何清洗步骤即可通过显微镜和流式细胞仪监测荧光信号。研究了探针长度，修饰骨架，探针浓度和细菌生长状态的影响。数据表明，基于16S rRNA的单核苷酸差异，三种密切相关的细菌（大肠杆菌，沙门氏菌和恶臭假单胞菌）之间存在特异性荧光鉴别。在对数中期或滞后阶段可以区分细胞。这些结果表明，QUAL探针可用于快速鉴定实验室和临床环境中的微生物。

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