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利用 DNA 微阵列检测和鉴定与水产品相关的细菌病原体。

Use of a DNA microarray for detection and identification of bacterial pathogens associated with fishery products.

机构信息

TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 HongDa Street, TEDA, Tianjin 300457, China.

出版信息

Appl Environ Microbiol. 2011 Dec;77(23):8219-25. doi: 10.1128/AEM.05914-11. Epub 2011 Sep 30.

Abstract

We established a microarray for the simultaneous detection and identification of diverse putative pathogens often associated with fishery products by targeting specific genes of Listeria monocytogenes, Salmonella, Shigella, Staphylococcus aureus, Streptococcus pyogenes, Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, and Yersinia enterocolitica and the 16S-23S rRNA gene internal transcribed spacer (ITS) region of Proteus mirabilis and Proteus vulgaris. The microarray contained 26 specific probes and was tested against a total of 123 target bacterial strains that included 55 representative strains, 68 clinical isolates, and 45 strains of other bacterial species that belonged to 8 genera and 34 species, and it was shown to be specific and reproducible. A detection sensitivity of 10 ng DNA or 10 CFU/ml for pure cultures of each target organism demonstrated that the assay was highly sensitive and reproducible. Mock and real fishery product samples were tested by the microarray, and the accuracy was 100%. The DNA microarray method described in this communication is specific, sensitive, and reliable and has several advantages over traditional methods of bacterial culture and antiserum agglutination assays.

摘要

我们建立了一个微阵列,通过针对李斯特菌、沙门氏菌、志贺氏菌、金黄色葡萄球菌、化脓性链球菌、霍乱弧菌、副溶血性弧菌、创伤弧菌和肠炎耶尔森菌的特定基因以及变形杆菌和普通变形杆菌的 16S-23S rRNA 基因内转录间隔区(ITS)区域,同时检测和鉴定与渔业产品相关的多种潜在病原体。该微阵列包含 26 个特定探针,经过 55 个代表性菌株、68 个临床分离株和 45 个其他细菌物种的总共 123 个目标细菌菌株的测试,结果表明该微阵列具有特异性和可重复性。每个目标生物的纯培养物的 10ng DNA 或 10CFU/ml 的检测灵敏度表明该检测方法非常灵敏且可重现。通过微阵列测试了模拟和真实的渔业产品样本,准确率为 100%。与传统的细菌培养和抗血清凝集检测方法相比,本研究中描述的 DNA 微阵列方法具有特异性、敏感性和可靠性等优点。

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16S-23S rDNA internal transcribed spacer regions in four Proteus species.四种变形杆菌属细菌的16S-23S核糖体DNA内转录间隔区
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