High-throughput suspension array for detecting four pathogens
High-throughput suspension array for detecting four pathogens
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用于检测四种病原体的高通量悬浮芯片
DOI：
10.1039/c2ay25327e
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发表时间：
2012-07
影响因子：
2
通讯作者：
Zhixian Gao
中科院分区：
文献类型：
--
作者：
Zhiyong Sun;Baoan Ning;Pu Su;Yuan Peng;Jialei Bai;Huishuang Chen;Jianqing Liu;Zhixian Gao
提出了一种同时定量检测食品中四种病原菌的悬浮阵列技术。根据从GenBank下载的大肠埃希菌、金黄色葡萄球菌、副溶血性弧菌和沙门氏菌的16S rDNA基因序列，设计了4组引物和种特异性捕获探针。将特定的核酸探针共价结合到荧光微球表面，用于液体悬浮杂交。将从样品中获得的生物素标记的PCR产物与其互补的核酸探针杂交，并在添加链霉亲和素藻红蛋白后进行检测。产物间无交叉反应，单通道法的灵敏度为5×10−6 mm。在多通道方法中，对大肠杆菌和沙门氏菌的灵敏度为5×10−4 mM，对金黄色葡萄球菌和副溶血性弧菌的灵敏度为5×10−5 mM。该多通道检测方法实现了多种病原体的多重检测。对于实际样品的检测，该方法在37℃下浓缩24小时后，对每克蔬菜的灵敏度为103CFU/g。这项研究证明了针对16S rDNA基因的悬浮阵列在检测食品样品中是否存在这四种病原体方面的有效性。
A new suspension array technology is proposed for the simultaneous quantitative determination of four pathogens in food. Four sets of primers and species-specific capture probes were designed based on the 16S rDNA gene sequences of Escherichia coli, Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella downloaded from GenBank. The specific nucleic acid probes were covalently bound to the surface of fluorescent microspheres for liquid suspension hybridization. The biotin-labeled PCR products obtained from the samples were hybridized with their complementary nucleic acid probes, which were detected after the addition of streptavidin phycoerythrin. No cross-reaction occurred among the products, and the sensitivity of the probe in the single-channel method was 5 × 10−6 mM. In the multi-channel method, the sensitivities were as follows: 5 × 10−4 mM for E. coli and Salmonella, and 5 × 10−5 mM for S. aureus and V. parahaemolyticus. The multi-channel method achieved multidetection of several pathogens. For real sample detection, the methods showed a sensitivity of 103 CFU per g to 100 CFU per g of vegetables after enrichment for 24 h at 37 °C. This study demonstrates the utility of the suspension array specific for the 16S rDNA gene for determining the presence of the four pathogens in food samples.
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影响因子：
3
作者：
Ming-Xia Yang;Han-Bo Zhang
通讯作者：
Ming-Xia Yang;Han-Bo Zhang
影响因子：
4.9
作者：
K. Yokogawa;S. Kawata;S. Nishimura;Y. Ikeda;Y. Yoshimura
通讯作者：
K. Yokogawa;S. Kawata;S. Nishimura;Y. Ikeda;Y. Yoshimura
影响因子：
3.3
作者：
W. Wilson;A. Erler;S. Nasarabadi;E. Skowronski;Paula M. Imbro
通讯作者：
W. Wilson;A. Erler;S. Nasarabadi;E. Skowronski;Paula M. Imbro
影响因子：
4.4
作者：
Nishigaki Y;Ueno H;Coku J;Koga Y;Fujii T;Sahashi K;Nakano K;Yoneda M;Nonaka M;Tang L;Liou CW;Paquis-Flucklinger V;Harigaya Y;Ibi T;Goto Y;Hosoya H;DiMauro S;Hirano M;Tanaka M
通讯作者：
Tanaka M
影响因子：
9.4
作者：
D. Egger;L. Pasamontes;Marianne Ostermayer;K. Bienz
通讯作者：
D. Egger;L. Pasamontes;Marianne Ostermayer;K. Bienz