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基于聚糖的流感病毒高特异性快速安培检测法

Highly specific and rapid glycan based amperometric detection of influenza viruses.

作者信息

Cui Xikai, Das Amrita, Dhawane Abasaheb N, Sweeney Joyce, Zhang Xiaohu, Chivukula Vasanta, Iyer Suri S

机构信息

788 Petit Science Center , Department of Chemistry , Center for Diagnostics and Therapeutics , Georgia State University , Atlanta , GA 30302 , USA . Email:

Atlanta Metropolitan State College , 1630 Metropolitan Parkway , Atlanta , GA 30310 , USA.

出版信息

Chem Sci. 2017 May 1;8(5):3628-3634. doi: 10.1039/c6sc03720h. Epub 2017 Feb 14.

Abstract

Rapid and precise detection of influenza viruses in a point of care setting is critical for applying appropriate countermeasures. Current methods such as nucleic acid or antibody based techniques are expensive or suffer from low sensitivity, respectively. We have developed an assay that uses glucose test strips and a handheld potentiostat to detect the influenza virus with high specificity. Influenza surface glycoprotein neuraminidase (NA), but not bacterial NA, cleaved galactose bearing substrates, 4,7di-OMe -acetylneuraminic acid attached to the 3 or 6 position of galactose, to release galactose. In contrast, viral and bacterial NA cleaved the natural substrate, -acetylneuraminic acid attached to the 3 or 6 position of galactose. The released galactose was detected amperometrically using a handheld potentiostat and dehydrogenase bearing glucose test strips. The specificity for influenza was confirmed using influenza strains and different respiratory pathogens that include and ; bacteria do not cleave these molecules. The assay was also used to detect co-infections caused by influenza and bacterial NA. Viral drug susceptibility and testing with human clinical samples was successful in 15 minutes, indicating that this assay could be used to rapidly detect influenza viruses at primary care or resource poor settings using ubiquitous glucose meters.

摘要

在即时检测环境中快速、准确地检测流感病毒对于采取适当的应对措施至关重要。目前的方法,如基于核酸或抗体的技术,分别存在成本高或灵敏度低的问题。我们开发了一种检测方法,该方法使用血糖仪试纸和手持式恒电位仪来高特异性地检测流感病毒。流感病毒表面糖蛋白神经氨酸酶(NA),而非细菌NA,可切割带有半乳糖的底物,即连接在半乳糖3或6位的4,7-二-O-甲基-乙酰神经氨酸,以释放半乳糖。相比之下,病毒和细菌的NA可切割天然底物,即连接在半乳糖3或6位的N-乙酰神经氨酸。使用手持式恒电位仪和带有脱氢酶的血糖仪试纸通过安培法检测释放的半乳糖。使用流感毒株和包括[具体细菌名称1]和[具体细菌名称2]在内的不同呼吸道病原体证实了对流感的特异性;细菌不会切割这些分子。该检测方法还用于检测由流感和细菌NA引起的合并感染。在15分钟内成功完成了病毒药物敏感性检测以及对人类临床样本的检测,这表明该检测方法可用于在基层医疗或资源匮乏的环境中使用普遍存在的血糖仪快速检测流感病毒。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32ac/5437373/a37215491034/c6sc03720h-f1.jpg

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